bluuebunny/crossref_metadata_2025 huggingface dataset:
REUTILIZATION OF LYMPHOCYTE DNA BY CELLS OF INTESTINAL CRYPTS AND REGENERATING LIVER
Authors: Bryant B. J.
Published: May 2004

Lymphoid cells from mice injected 54 hours and 30 hours earlier with 3H-thymidine were washed and transfused into isogenic recipients at 29 to 30 hours after partial hepatectomy. The recipients were killed 28 to 30 hours later, and liver, intestine, Peyer's patch, spleen, and the transfused cells were examined in autoradiographs exposed 6 months. Approximately 80 per cent of the labeled transfused cells were classed as lymphocytes. The labeled DNA contained in the transfused cells was partitioned to about 14 times as many recipient liver and intestinal cells, appearing in 72 to 78 per cent of hepatocyte nuclei, in 30 to 35 per cent of liver reticuloendothelial nuclei, and in 90 to 95 per cent of intestinal crypt nuclei. The label was not comparably widespread in the lymphoid organs, but was limited to a few intensely labeled lymphocytes and a somewhat larger number of very weakly labeled cells. When heat-killed cells rather than living cells were transfused, intensely labeled lymphocytes were absent from the lymphoid organs, but the labeling of cells in the recipients was otherwise identical. The results suggest that (a) reutilized DNA is derived from dead cells, (b) reutilized DNA is mainly degraded to nucleosides and nucleotides, the usual immediate de novo DNA precursors, before reincorporation into DNA, and (c) DNA reutilization may occur in the lymphoid organs, but on a less active scale than in intestine or regenerating liver.
FINE STRUCTURAL CHANGES IN THE INTESTINAL EPITHELIUM OF THE BULLFROG DURING METAMORPHOSIS
Authors: Bonneville Mary A.
Published: May 2004

The fine structural changes occurring in the columnar absorbing cells of the intestinal epithelium during metamorphosis of the bullfrog, Rana catesbeiana, have been examined by phase contrast and electron microscopy. Tissue samples taken just posterior to the entrance of the hepatopancreatic duct were fixed in veronal acetate-buffered osmium tetroxide and embedded in methacrylate. Under the action of the metamorphic stimulus (thyroid hormone), specific and characteristic responses were given by differentiated larval cells and undifferentiated basal cells within the same epithelium. The functional larval cells underwent degenerative changes and were retained for a time within the metamorphosing epithelium. Dense bodies appeared and increased in number in association with the loss of normal cell structure. Because of their morphology and time of formation, these bodies have been tentatively identified as lysosomes. Early in metamorphosis the basal cells did not change, but they subsequently proliferated to form a new cell layer beneath the remaining degenerating cells that lined the lumen. After the dying cells were sloughed into the gut, the new epithelium differentiated to form the adult tissue. The columnar epithelial cells of the mature animal differed in their fine structural organization from their larval precursors. Therefore, their adult configuration was molded by the action of the metamorphic stimulus.
RELATION OF PROTEIN SYNTHESIS TO THE DIVISION CYCLE IN MAMMALIAN CELL CULTURES
Authors: Taylor Edwin W.
Published: May 2004

Protein synthesis in suspension cultures of human cells in logarithmic growth was inhibited with puromycin or chloramphenicol, and the growth rate and mitotic index were measured as a function of time. The mitotic index remained constant for about 1 hour after addition of inhibitor; this indicates that any protein synthesis necessary for mitosis is completed before the beginning of prophase. For rates of protein synthesis equal to or greater than 0.3 that of untreated cells, the index decreased over a 5- to-7-hour period and then remained constant. The final value of the index relative to that of the uninhibited control was approximately equal to the relative rate of protein synthesis. The period from the end of DNA synthesis to mitosis (G2) was increased by partial inhibition of protein synthesis. A mathematical model of the inhibition process has been formulated which predicts the shape of the mitotic index curves and the increase in the G2 period. An interpretation of the model is that the rate-limiting step is the synthesis of an enzyme which catalyzes the formation of a compound necessary to initiate mitosis.
STUDIES ON THE ISOLATION OF METAPHASE CHROMOSOMES
Authors: Choraży M. , Bendich A. , Borenfreund E. , Hutchison D. J.
Published: May 2004

A method for the isolation of metaphase chromosomes from mouse L1210 leukemia cells has been developed. Cells, arrested at metaphase with colchicine, were exposed to hypotonic solution and the pH was then adjusted to 5.6 to stabilize the chromosomes. The metaphase figures were subsequently disrupted and the chromosomes isolated by a series of differential centrifugations in sucrose. The isolated chromosomes were well preserved, as judged by morphological criteria. The effect of various enzymes and chemical agents on the isolated chromosomes was studied. Chymotrypsin, trypsin, and deoxyribonuclease caused a marked disintegration of the chromosomes, whereas treatment with pepsin and ribonuclease induced no significant morphological alterations.
THE BASE COMPOSITION OF RIBONUCLEIC ACID IN LAMPBRUSH CHROMOSOMES, NUCLEOLI, NUCLEAR SAP, AND CYTOPLASM OF TRITURUS OOCYTES
Authors: Edström J. -E. , Gall J. G.
Published: May 2004

The base composition of RNA's extracted from chromosomes, nucleoli, nuclear sap, and cytoplasm of Triturus oocytes has been determined by microelectrophoresis. The chromosomal RNA has a content of guanine+cytosine equal to that of DNA, but there is no complementarity in the composition as for DNA. Nuclear sap contains a highly variable RNA with a tendency towards high uracil values. Nucleolar and cytoplasmic RNA's are similar in composition and both are of the guanine-cytosine rich type. The chromosomes and nucleoli contain roughly equivalent amounts of RNA, somewhat less than is present in the nuclear sap. The RNA/DNA ratio of the whole chromosomes is about 10. However, the ratio in the synthetically active regions, the loops, is much higher, since the loops contain all the chromosomal RNA but only a small fraction of the DNA.
ISOLATION OF AN INSULIN SECRETION GRANULE FRACTION
Authors: Lindall Arnold W. , Bauer G. Eric , Dixit Padmaker K. , Lazarow Arnold
Published: May 2004

Goosefish islets were homogenized in 0.25 M sucrose and separated into nuclear, mitochondrial + secretion granule, microsomal, and supernatant fractions. Eighty per cent of the cytochrome oxidase activity and 75 per cent of the bioassayed insulin activity were found in the mitochondrial + secretion granule fraction (6000 g for 10 minutes). The mitochondrial + secretion granule fraction was further subfractionated by centrifugation (2 hours at 100,000 g and 0°C) using a continuous linear density gradient 1.0–2.0 M sucrose). Eighteen to 20 subfractions were collected by piercing the bottom of the tube and collecting drops. The total protein was distributed into a bimodal curve consisting of a high density component, which contained 90 per cent of the insulin (secretion granules), and a lower density component, which contained the cytochrome oxidase activity (mitochondria).
ELECTRON MICROSCOPE OBSERVATIONS ON THE SURFACE ADENOSINE TRIPHOSPHATASE-LIKE ENZYMES OF HELA CELLS INFECTED WITH HERPES VIRUS
Authors: Epstein M. A. , Holt S. J.
Published: May 2004

HeLa cells infected with herpes simplex virus have been examined in thin sections by electron microscopy after cytochemical staining for the presence of surface enzymes splitting adenosine triphosphate. As with uninfected HeLa cultures (18), the opaque enzyme reaction product was localized at the plasma membranes of about half the cells, tending to be present where there were microvilli and absent on smooth surfaces. Where mature extracellular herpes particles were found in association with cell membranes showing the enzyme activity, they were invariably likewise stained, and conversely, those mature particles which lay close against cells without reaction product at the surface were themselves free of it. Particles found budding into cytoplasmic vacuoles were also always without opaque deposit since this was never seen at vacuolar membranes, even in cells having the activity at the surface. The enzyme reaction product thus provided a marker indicating the manner in which the particles escape from cells and mature by budding out through cellular membranes, carrying, in the process, a portion of the latter on to themselves to form the outer viral limiting membrane. In some instances, virus particles were observed with more opaque material covering them than was present at the cell membrane with which they were associated. This finding has been taken as evidence for a physiological waxing and waning of surface enzyme activity of adenosine triphosphatase type. The fine structure of the mature extracellular virus as prepared here, using glutaraldehyde fixation, is also recorded. The observations and interpretations are discussed in full.
CHOLINESTERASE IN DENERVATED END PLATES AND MUSCLE FIBRES
Authors: Brzin Miro , Majcen-Tkačev Živa
Published: May 2004

Parallel studies were made of cholinesterase activities and localizations in denervated rat and rabbit gastrocnemius muscle. Koelle's histochemical reaction was used for demonstrating the localization of cholinesterases. Enzyme activities in whole sliced muscle were measured by electrometric titration. The Cartesian ampulla-diver technique was used for cholinesterase activity determinations in end plate regions or in small pieces of the muscle fibre itself. No changes in the activity of cholinesterases (ChE) were found in the whole denervated muscle which would account for its chemical supersensitivity. The ChE distribution pattern was changed so that the end plate region became less active in the denervated muscle than in the normal one. The decrease in ChE activity in the end plates seems to be largely compensated for by an increase of this enzyme elsewhere in the muscle. A possible connection between the spatial spread of cholinesterase activity and the enlargement of the acetylcholine-sensitive surface is discussed.
CHEMICAL DIFFERENTIATION OF NUCLEAR PROTEINS DURING SPERMATOGENESIS IN THE SALMON
Authors: Alfert Max
Published: May 2004

By means of qualitative staining experiments the characteristic protein changes which occur during maturation of salmon sperm can be followed. It can be observed that the replacement of histone by protamine takes place after completion of meiosis during an advanced stage of spermiogenesis.
ALTERATIONS IN BIOCHEMICAL COMPOSITION AND RIBONUCLEIC ACID METABOLISM INDUCED IN RAT LIVER BY CORTISONE
Authors: Lowe Charles Upton , Rand Royden N.
Published: May 2004

An investigation of the effect of cortisone administration upon the chemical composition of intracellular particulates of rat liver has been made. Livers were homogenized in 0.25 M sucrose solutions and submitted to differential centrifugation. Five fractions were prepared: mitochondria (Mit), microsomes (Mi), ultracentrifugable (U), non-sedimentable (S), and nuclear (Nuc). Measurement was made of total and polymerized RNA, nitrogen, lipide P, and uptake of P32 by the RNA of each fraction. The following observations were made:—

Cortisone administration caused a fall in concentration in all measured constituents except glycogen. On a per liver basis, however, total liver RNA was unchanged in amount; nitrogen content of Mi fell and that of S increased; the lipide P of Mit and Mi also decreased. The biochemical composition of a statistical mitochondrion was significantly altered; in contrast, the microsomal fraction decreased in amount, but the relationship between the chemical constituents was unchanged.

When polymerized RNA was sought by a process involving precipitation from ethanol at 20°C., none was found in the Mit of cortisone livers and the amount in Mi was much less than found in the normal. When, however, precipitation was conducted at 4°C., yields of polymerized RNA in all fractions after cortisone were equal to or greater than those found in the normal. Furthermore, incubation of mixtures of homogenates from normal and cortisone livers resulted in loss of warm precipitable RNA.

These data strongly suggest the presence of an enzyme in cortisone livers which upon incubation with normal livers made preparation of polymerized RNA virtually impossible by use of the warm method. This agent, thought to operate in vivo and in vitro, was not present in significant amounts in normal livers, since incubation in this instance had no effect upon the amount of polymerized RNA.

Mit from cortisone livers obtained by the cold technique had a significantly decreased rate of incorporation of P32 even though the yield of RNA from this fraction was increased. To reconcile these observations, it was proposed that under the influence of cortisone a variant of normal RNA is synthesized or normal RNA is converted to this variant. This "new" RNA has new solubility properties, a new rate of incorporation of P32, and conceivably it cannot act as a template for normal protein synthesis.
ELECTRON MICROSCOPE STUDIES ON THE DICTYOSOMES AND ACROBLASTS IN THE MALE GERM CELLS OF THE CRICKET
Authors: Beams H. W. , Tahmisian T. N. , Devine R. L. , Anderson Everett
Published: May 2004

The dictyosome (Golgi body) in the secondary spermatocyte of the cricket appears in electron micrographs as a duplex structure composed of (a) a group of parallel double-membraned lamellae and (b) a group of associated vacuoles arranged along the compact lamellae in a chain-like fashion. This arrangement of ultramicroscopic structure for the dictyosomes is strikingly comparable to that described for the Golgi apparatus of vertebrates. Accordingly, the two are considered homologous structures. Associated with the duplex structure of the dictyosomes is a differentiated region composed of small vacuoles. This is thought to represent the pro-acrosome region described in light microscope preparations.

In the spermatid the dictyosomes fuse, giving rise to the acroblast. Like the dictyosomes, the acroblasts are made up of double-membraned lamellae and associated vacuoles. In addition, a differentiated acrosome region is present which, in some preparations, may display the acrosome vacuole and granule.

Both the dictyosomes and acroblasts are distinct from mitochondria.
VARIATIONS IN THE STRUCTURE OF MITOCHONDRIA
Authors: Dempsey E. W.
Published: May 2004

A characteristic internal structure, consisting of a double-layered outer wall enclosing a matrix-filled space through which pass double-layered membranous folds, would appear to comprise as satisfactory a definition of mitochondria for electron microscopy as their intravital affinity for Janus green affords for light microscopy. Relying for identification upon this characteristic internal structure, mitochondria appear to be pleomorphic structures which vary in size, shape, complexity, and density. They are labile also in that their number may increase or decrease under controlled conditions. The possibility therefore exists that these organelles are constantly being formed and destroyed, perhaps by their participation in metabolic processes.

The problem of the origin of mitochondria is in an unsatisfactory state. New organelles unquestionably are formed in particular physiological states. The possibility that new bodies are produced by fission of ones already present does not seem adequate. On the other hand, the possible fabrication of new mitochondria out of intracellular membranes, although an attractive hypothesis, has not been adequately substantiated.
STUDIES ON THE CROSS-STRIATION OF THE INDIRECT FLIGHT MYOFIBRILS OF THE BLOWFLY CALLIPHORA
Authors: Hanson Jean
Published: May 2004

1. The cross-striation in the indirect flight myofibrils of Calliphora has been studied by phase contrast and polarised light microscopy. The band pattern at rest-length has been determined in flies killed in osmium tetroxide vapour while their wings remained in the resting position. All other observations have been made on unfixed fibrils. Although length changes in situ are probably very slight (about 2 per cent), isolated fibrils, by treatment with crude muscle extract or with ATP, can be induced to elongate to 104 per cent rest-length, or to shorten by 8 per cent but no more. Over the range 98 to 104 per cent rest-length, experimentally induced length changes are reversible. The fibrils can also be stretched beyond 104 per cent rest-length, but the process is irreversible. During the course of glycerol extraction the fibrils elongate to 104 per cent rest-length.

2. The changes in band pattern observed over the range 104 to 92 per cent rest-length are qualitatively the same as the changes observed over a wider range (about 130 to 40 per cent rest-length) in the skeletal myofibrils of rabbits. The earlier stages of shortening appear to be effected by retraction of the I bands into the A bands where they fill up the H zones. No evidence has been found that any changes in band pattern are due to a migration of the A substance.

3. Two components of the sarcomere can be extracted from it and a third component remains behind. These three components, which have also been demonstrated in skeletal myofibrils of the rabbit, where they behave in the same way, are: (a) the A substance which does not change its position as the fibril changes its length, and which can be extracted by the same procedures as remove myosin (shown elsewhere to be the A substance) from rabbit fibrils; (b) a material which extends from the Z lines to the borders of the H zone and which moves inwards during contraction and outwards during elongation; it can capture rabbit myosin from solution and form with it a contractile system, and it is thought to be actin; (c) a "backbone" or stroma bearing Z and M lines.

4. Since all these features of the cross-striation are the same in the insect fibrils as in rabbit fibrils, it is considered very probable that the sarcomere is similarly organised in both types of muscle and contracts by essentially the same mechanism.
AN ELECTRON MICROSCOPE STUDY OF THE ENDOPLASMIC RETICULUM IN NEWT NOTOCHORD CELLS AFTER DISTURBANCE WITH ULTRASONIC TREATMENT AND SUBSEQUENT REGENERATION
Authors: Selman G. G. , Jurand A.
Published: May 2004

Ultrasonic treatment of the tails of Triturus alpestris tadpoles, at intensities of 8 to 15 watts/cm2, at 1 megacycle/sec., for 5 minutes, disrupted the epidermis and caused pycnosis in individual cells of the muscle and neural tube, but caused no damage to the notochord that could be detected by light microscopy. Electron microscopy showed that this ultrasonic treatment disordered nearly all the endoplasmic reticulum (ER) of the notochord cells into irregularly rounded vesicles, but within 3 hours after treatment some parallel arrays of normal endoplasmic reticulum were seen near, and continuous with, the outer nuclear membrane. In addition, a re-ordering of the previously disordered ER took place throughout the cytoplasm, in some cases. A classification was made of the state of the ER as shown in electron micrographs of material fixed immediately, 3, and 24 hours after treatment. This showed that more than half the total endoplasmic reticulum in notochord cells was normal again by 24 hours after treatment.
FINE STRUCTURE OF THE NEUROHYPOPHYSIS OF THE OPOSSUM (DIDELPHIS VIRGINIANA)
Authors: Roth Lawrence M. , Luse Sarah A.
Published: May 2004

The neurohypophysis of the opossum (Didelphis virginiana) was studied by electron microscopy in order to amplify Bodian's classic light microscopic observations in which he demonstrated a definite lobular pattern. The lobule of the opossum neurohypophysis is divided into three regions: a hilar, a palisade, and a septal zone. The hilar portion contains bundles of nerve fibers, the extensions of the hypothalamo-hypophyseal tract containing neurofilaments but few neurosecretory granules. In the opossum, pituicytes have a densely fibrillar cytoplasm. Herring bodies are prominent in the hilar region. They are large bodies packed with neurosecretory granules that have been described as end bulb formations of axons. From the hilar region, axons fan out into a palisade zone where the nerve terminals packed with neurosecretory granules, mitochondria, and microvesicles abut upon basement membranes. The neurosecretory granules are similar to those present in the neurohypophysis of other mammals, except for an occasional huge granule of distinctive type. Material morphologically and histochemically resembling glycogen occurs as scattered particles and as aggregates within nerve fibers. The septal zone, containing collagen, fibroblasts, and numerous small capillaries, is separated from the adjacent glandular tissue by a basement membrane.
HYDROLYTIC ENZYMES OF RABBIT MONONUCLEAR EXUDATE CELLS
Authors: Dannenberg Arthur M. , Bennett William E.
Published: May 2004

Oil-induced mononuclear phagocytes (MN) were quantitatively assayed for various hydrolases as unfractionated suspensions of frozen and thawed cells. They apparently contain two proteases. The first, measured with urea- or acid-denatured hemoglobin, was similar to purified Proteinase I of lung with respect to pH optimum (pH 4), stability, hydrolytic and polymerizing activities, and reactions to various inhibitors. The second protease resembled chymotrypsin in its hydrolysis of glycyl-L-phenylalanine amide, acetyl-L-tyrosine ethyl ester and N-benzoyl-DL-phenylalanine-ß-naphthol ester (BPN). With the latter, its pH optimum was between 5.0 and 5.8, and its action was inhibited by diisopropylphosphorofluoridate (DFP) and p-chloromercuribenzoate. When assayed under the above conditions, polymorphonuclear exudate cells (PMN) and red blood corpuscles (RBC) manifested little or no hydrolysis of either hemoglobin or BPN. MN also contained esterases that split methyl butyrate and ß-naphthyl acetate. The pH optimum with the latter was 7.4, and its hydrolysis was partially inhibited by DFP, fluoride, taurocholate, and eserine. PMN had low esterase activity; RBC had little or none. MN, but not PMN or RBC, contained a stable lipase with a pH optimum of 6.1 in maleate buffer. Protamine, NaCl, heat, p-chloromercuribenzoate, ethylenediamine tetraacetate, taurocholate, and DFP were inhibitory, but no appreciable activation occurred in the presence of heparin or serum. Thus it possessed some of the characteristics of Korn's lipoprotein lipase, but not others.
FINE STRUCTURE OF CELL SURFACE SPECIALIZATIONS IN THE MATURING DUODENAL MUCOSA OF THE CHICK
Authors: Overton Jane , Shoup Jane
Published: May 2004

Cell surfaces in the duodenal mucosa have been studied in maturing tissue of the chick from incubation until hatching. Changes in the distribution of mitoses in this tissue give an indication of its rate of maturation. This rate is paralleled in developmental changes in microvilli and junctional complexes. Concentration of mitotic figures towards the base of villous folds is gradual from day 9 to day 16, then rapid to day 19, after which the mature pattern is acquired. By day 11, microvilli appear in a regular pattern which does not alter through hatching. Their height remains the same to day 16 when it increases gradually to day 19, then very sharply. The angle formed between the microvilli and the cell surface increases gradually to day 16, giving evidence of advancing internal structure. Changes in cell adhesion also occur at day 16. Thereafter, following trypsin treatment cells are held together in patches by the tight junctions of the terminal bar, although the desmosomes are separated. The timing of these morphological changes is compared with that of alkaline phosphatase accumulation in this tissue as reported by Moog (13). Increase in the surface area of the microvilli parallels the increase in the activity of the enzyme.
CYTOCHEMICAL DIFFERENCES IN KIDNEYS FROM WINTER-HIBERNATING AND AROUSED BATS (MYOTIS LUCIFUGUS), WITH PARTICULAR REFERENCE TO THE GOLGI ZONE
Authors: Rosenbaum Robert M. , Melman Arnold
Published: May 2004

Kidneys from winter bats (Myotis lucifugus) were removed and fixed in cold formalin-calcium while the animals were in the following states: (a) natural hibernation; (b) arousal from hibernation for 24 hours; (c) laboratory maintained hibernation; and (d) no hibernation since the previous winter. With fixed frozen sections, the lead salt method of Wachstein and Meisel with adenosine triphosphate as substrate (pH 7.2) showed enzymic activity localized in large vacuoles and smaller vesicles or droplets in the Golgi region of distal and proximal tubular epithelial cells of kidneys from hibernating bats. No ATPase activity was detected in the basal lamellae of tubular epithelium from hibernating bats. ATPase activity in the Golgi region was not seen in cells from kidney tubules of bats aroused from hibernation 24 hours previously or of animals that had not hibernated, whereas activity for ATPase was present in the basal infoldings of tubular epithelium from these animals. Inosine di- and triphosphatase and calcium activated ATPase activities were also detected in the Golgi region of hibernating bats but were not present in the basal infoldings of tubular epithelium from active animals. There was little or no activity toward the mono- and diphosphates of adenine, thiamine pyrophosphate, and the di- or triphosphates of guanidine, cytidine, or deoxyadenosine. The loss of enzymic activity from the Golgi region of the tubular epithelium from hibernating bats and its increase in the region of the basal infoldings of tubular epithelium in aroused bats suggests that the Golgi region plays a role in the synthesis of enzymic protein usually identified with the external cell membrane.
STUDIES ON LYSOSOMES
Authors: Weissmann Gerald , Becher Beverly , Thomas Lewis
Published: May 2004

Granules from rabbit peritoneal leucocytes were prepared in 0.3 M sucrose as an optically homogeneous suspension with the aid of heparin. Lysis of the granules in vitro was followed by measurement of decreases in the apparent absorbance of the suspensions at 520 mµ and was accompanied by solubilization of beta-glucuronidase from the particles. Streptolysins O and S from hemolytic streptococci lysed the granules at 20°C; the initial rate of lysis by streptolysin O was greater than that by streptolysin S. Cysteine activated, and specific antibody inhibited, streptolysin O; antimycin and bovine serum albumin inhibited streptolysin S. The granules were not lysed by any other streptococcal exotoxins. Lysis was irreversible and depended neither upon oxidative phosphorylation, nor upon intact respiration. The granules were also lysed by lysolecithin, at concentrations from 2 x 10-6 M to 1 x 10-4 M; bovine serum albumin and antimycin also inhibited this lytic agent. Such other hemolytic agents and procedures as vitamin A, non-ionic detergents, and ultraviolet irradiation also disrupted leucocyte granules. In susceptibility to lysis and other properties, the granules of white cells resembled erythrocytes. Leucocyte granules differed from mitochondria in that they did not appear to take up or extrude water reversibly; they were unaffected by thyroxine, phosphate, or metabolic substrate. The studies are compatible with the hypotheses that white cell granules are similar to lysosomes isolated from other tissues, and that they share common surface properties with erythrocytes.
THE FINE STRUCTURE OF GREEN BACTERIA
Authors: Cohen-Bazire Germaine , Pfennig Norbert , Kunisawa Riyo
Published: May 2004

The fine structure of several strains of green bacteria belonging to the genus Chlorobium has been studied in thin sections with the electron microscope. In addition to having general cytological features typical of Gram-negative bacteria, the cells of these organisms always contain membranous mesosomal elements, connected with the cytoplasmic membrane, and an elaborate system of isolated cortical vesicles, some 300 to 400 A wide and 1000 to 1500 A long. The latter structures, chlorobium vesicles, have been isolated in a partly purified state by differential centrifugation of cell-free extracts. They are associated with a centrifugal fraction that has a very high specific chlorophyll content. In all probability, therefore, the chlorobium vesicles are the site of the photosynthetic apparatus of green bacteria.
AN ANALYSIS OF COLLAGEN SECRETION BY ESTABLISHED MOUSE FIBROBLAST LINES
Authors: Goldberg Burton , Green Howard
Published: May 2004

In vitro synthesis of collagen by established mouse fibroblast lines has been examined by electron microscopy. During rapid growth (log phase), when collagen could not be detected in the cultures, the cells lacked a well developed granular ergastoplasm and Golgi system. Upon cessation of growth (stationary phase), collagen accumulated in the cultures and the cells demonstrated highly developed granular and smooth ergastoplasm. Collagen appeared to be synthesized in the rough-surfaced endoplasmic reticulum and to be transported as a soluble protein to the cell surface by vesicular elements of the agranular ergastoplasm. Fusion of the limiting membranes of these vesicles with the cell membrane permitted the discharge of the soluble collagen into the extracellular space, where fibrils of two diameter distributions formed. The secretion of collagen is concluded to be of the merocrine type. Alternative theories of collagen secretion are discussed and the data for established lines compared with the results of other in vitro and in vivo studies of collagen fibrillogenesis.
INCORPORATION OF TRITIATED THYMIDINE IN THE CELLS OF CAENORHABDITIS BRIGGSAE (NEMATODA) REARED IN AXENIC CULTURE
Authors: Nonnenmacher-Godet Jacqueline , Dougherty Ellsworth C.
Published: May 2004

In the rhabditid nematode Caenorhabditis briggsae the incorporation of thymidine-H3 has been studied by autoradiography after Feulgen staining, with animals maintained under axenic conditions in a medium of only partly defined composition. Labeling has been followed in adults left in the presence of thymidine-H3 for periods of from ½ to 24 hours, as well as in adults reared from larvae in the presence of the tritiated nucleoside. A massive incorporation is found in the nuclei of the gonads and intestine; also a less intense particulate cytoplasmic incorporation is clear in certain cells, especially those of the intestine. In general, all labeling has proved to be sensitive to DNase, but resistant to RNase. The label's stability has been tested by the transfer of adults into a medium containing "cold" thymidine. They remain there for up to 48 hours. A transfer for 24 hours results in a considerable decrease in the intensity of nuclear and cytoplasmic labeling; a stay of 48 hours leads to its complete disappearance from non-dividing (intestinal) as well as dividing (gonadal) nuclei. A phenomenon of DNA turnover is envisaged and discussed as a possible physiological attribute of C. briggsae.
FINE STRUCTURAL CHANGES IN PROPLASTIDS DURING PHOTODESTRUCTION OF PIGMENTS
Authors: Klein Shimon , Bogorad Lawrence
Published: May 2004

Etiolated bean leaves supplied δ-amino-levulinic acid in the dark synthesize large amounts of protochlorophyllide which is not converted to chlorophyllide upon illumination of the leaves. The fine structure of the proplastids is not affected by the treatment. When leaves containing "inactive" protochlorophyllide are exposed to light of 700 ft-c for 3 hours, they lose practically all their green pigments. During this period large stacks of closed membrane structures are built up in the region of the prolamellar body. These lamellar structures remain even when no or only traces of pigment are left in the leaves. In untreated control leaves the pigment content remained constant during similar illumination and the structural changes in the plastids consisted of a rearrangement of the vesicles from the prolamellar bodies into strands dispersed through the stroma; lamellae and grana formation occurred later.
EFFECTS OF PUROMYCIN ON THE NUCLEOPROTEINS OF THE HELA CELL
Authors: Studzinski G. P. , Love R.
Published: May 2004

The effects of several concentrations of puromycin on the nucleoproteins of HeLa cells grown in monolayers were studied by cytochemical and biochemical techniques. The earliest change at all concentrations of puromycin was a decrease in a granular form of ribonucleoprotein (RNP) that is demonstrable in the normal HeLa cell by the toluidine blue-molybdate (TBM) stain. The other types of RNP revealed by the TBM method were unaltered although the cell volume decreased markedly. Treatment with high concentrations of the antimetabolite resulted in pre-prophase inhibition of mitotic division and led to production of inclusions containing RNP in the cytoplasm; lower concentrations resulted in metaphase arrest. Biochemical analyses confirmed the cytochemical observations and indicated that synthesis of RNA and protein was inhibited to the same extent.
MEASUREMENTS OF SOME CELLULAR CHANGES DURING THE FIXATION OF AMPHIBIAN ERYTHROCYTES WITH OSMIUM TETROXIDE SOLUTIONS
Authors: Tooze John
Published: May 2004

On average, 15 per cent of the total haemoglobin present in the blood of the newt Triturus cristatus was extracted during 45 minutes of fixation in Palade-Caulfield fixative. This extraction was reduced with fixatives buffered at pH 6.2 instead of pH 7.4. The addition of Ca++ ions to a final concentration of 0.01 M in the fixative completely suppressed haemoglobin extraction. The effect of the pH, and the presence or absence of Ca++ ions in the fixative, on the rate of haemoglobin extraction has been determined. During Palade-Caulfield fixation the average projected area of newt erythrocytes increased by 37 per cent, and after dehydration and embedding in Epon the average area was 25 per cent greater than that of the unfixed cell. Fixatives buffered at pH 6.2 and containing 0.01 M Ca++ ions caused cellular shrinkage, with the average projected area decreasing by 10 per cent in the fixative. This shrinkage continued during dehydration, and the final average area of the erythrocytes in Epon was 26 per cent less than that of the unfixed cells. Similar measurements with erythrocytes of Amphiuma tridactylum showed that after Palade-Caulfield fixation the average cellular area was increased by 45 per cent, and after dehydration and embedding in Araldite it was 36 per cent greater than that of the unfixed cell. The average nuclear area increased by 35 per cent during fixation but after embedding it was 26 per cent greater than that of the unfixed nuclei. With a fixative at pH 6.2 containing 0.01 M Ca++ ions, both the nucleus and the whole cell shrank during fixation. The nuclear area decreased by 20 per cent and the cellular area by 22 per cent. After dehydration and embedding in Araldite, the average nuclear area had decreased by 35 per cent and the cellular area by 40 per cent. It has been shown that OsO4 fixation lowers the isoelectric points of haemoglobins and other proteins. This finding has been used in the interpretation of the observed cellular changes resulting from fixation.
STUDIES ON AN EPITHELIAL (GLAND) CELL JUNCTION
Authors: Loewenstein Werner R. , Kanno Yoshinobu
Published: May 2004

Membrane permeability of an epithelial cell junction (Drosophila salivary gland) was examined with intracellular microelectrodes and with fluorescent tracers. In contrast to the non-junctional cell membrane surface, which has a low permeability to ions (10-4 mho/cm2), the junctional membrane surface is highly permeable. In fact, it introduces no substantial restriction to ion flow beyond that in the cytoplasm; the resistance through a chain of cells (150 Ω cm) is only slightly greater than in extruded cytoplasm (100 Ω cm). The diffusion resistance along the intercellular space to the exterior, on the other hand, is very high. Here, there exists an ion barrier of, at least, 104Ω cm2. As a result, small ions and fluorescein move rather freely from one cell to the next, but do not leak appreciably through the intercellular space to the exterior. The organ here, rather than the single cell, appears to be the unit of ion environment. The possible underlying structural aspects are discussed.
THE PREPARATION AND PROPERTIES OF BACTERIAL CHROMATOPHORE FRACTIONS
Authors: Worden Patricia Barron , Sistrom W. R.
Published: May 2004

Chromatophore material from the bacterium Rhodopseudomonas spheroides was freed of ribosomes by centrifugation in 27 per cent RbCl and then separated into "heavy" and "light" fractions by centrifugation through a sucrose gradient. The fractions differed from one another in the following ways. (a) The isopycnic density of the heavy fraction was between 1.15 and 1.18 gm/ml and that of the light fraction was 1.14 gm/ml. (b) The heavy fraction was able to bind ribosomes; the light fraction was not. (c) The light fraction was homogeneous in the ultracentrifuge and had a sedimentation constant, extrapolated to infinite dilution, of 153 s20,w. The heavy fraction was grossly heterogeneous. (d) Both the amount of bacteriochlorophyll relative to protein and the ratio of bacteriochlorophyll to carotenoids were greater in the light fraction. (e) The spectra of the two fractions in the near infra-red were different. Comparisons of the chromatophore fractions from cells with different amounts of bacteriochlorophyll showed that the specific bacteriochlorophyll contents of the two fractions did not change to the same extent as did that of the whole cells. The amount of heavy fraction from pigmented cells was roughly independent of the cellular pigment content and was about equal to that from pigment-free cells. The amount of light fraction depended on the pigment content of the cells; no light fraction was obtained from cells devoid of bacteriochlorophyll. The cytochrome complements of both fractions underwent quantitative as well as qualitative changes with varying growth conditions. The size of the photosynthetic unit in R. spheroides appeared to increase as the total cellular bacteriochlorophyll content increased; however, the number of units per light fraction particle remained constant.
A NUCLEAR MEMBRANE CHANGE AFTER PARTIAL HEPATECTOMY
Authors: Kishimoto Susumu , Lieberman Irving
Published: May 2004

Partial hepatectomy (67 per cent extirpation) of the rat leads to a change in the membrane of liver nuclei (purified with citric acid) detectable as an increase in electrophoretic mobility. No change is detectable 2 hours after the operation, but between 2 and 6 hours about a 1.4-fold increase in mobility occurs after which the mobility becomes constant at the elevated level. Removal of only 10 per cent of the liver causes no detectable change in 6 hours. Bilateral adrenalectomy immediately before partial hepatectomy does not affect the development of the nuclear change. Actinomycin D and p-fluorophenylalanine, but not noradrenalin, ionizing radiation, or EDTA, suppress the increase in electrophoretic mobility. The level of actinomycin D required to block the nuclear membrane change is 6 times greater than that necessary to prevent the rate increase in hepatic RNA metabolism that follows removal of part of the liver. This discrepancy and the difference in the response to noradrenalin indicate that, at least initially, the nuclear membrane change and the change in the rate of RNA synthesis are independent processes. The inability of EDTA to block the nuclear membrane change shows that the Zn++ requirement for DNA replication is not related to the events that lead to the alteration in the electrokinetic properties of liver nuclei.
EVIDENCE FOR CYTOPLASMIC DNA IN ROOT CELLS OF NICOTIANA
Authors: Mourad Effat Badr
Published: May 2004

Sterile root cultures from Nicotiana tabacum were grown with H3-thymidine added to the medium for various intervals. Incorporation of the labeled nucleoside into nuclear DNA occurred in a fraction of the nuclei which increased with time. In addition, the cytoplasm of all cells incorporated enough tritium to be readily detected by autoradiography. The tritium was not removed by hydrolysis in 1 N HCl at 60°C for 10 minutes, but was removed by digestion in a DNase solution which also removed nuclear DNA. The amount of tritium in the cytoplasm increased during the first 2 hours, but did not appear to increase significantly during the following 5 hours. If the roots were transferred to unlabeled medium after 2 hours, the label was diluted faster than expected by growth without turnover of the labeled component. If FUdR was added to the unlabeled medium, the depletion occurred faster during the first 6 hours, but later appeared to level off so that at 10 hours these cultures did not differ from those incubated without FUdR. However, the addition of an excess of unlabeled carrier had no effect on the rate of depletion of the cytoplasmic label. Actinomycin D, which inhibited the incorporation of H3-cytidine into RNA in the root tips, had no effect on the incorporation of H3-thymidine into the cytoplasmic component. However, Mitomycin C or a high concentration of deoxyadenosine inhibited the incorporation of H3-thymidine into the cytoplasmic component as well as into the nuclear DNA. It is concluded that H3-thymidine is incorporated into a cytoplasmic fraction which has the characteristics of DNA, with a measurable rate of turnover. This fraction is synthesized regardless of whether or not the nucleus is synthesizing DNA. Although the function of cytoplasmic fraction is not yet known, it does not appear to be that of supplying precursors for the synthesis of the nuclear DNA.
A NEW APICAL MICROTUBULE-ASSOCIATED ORGANELLE IN THE STERNAL GLAND OF ZOOTERMOPSIS NEVADENSIS (HAGEN), ISOPTERA
Authors: Satir P. , Stuart A. M.
Published: May 2004

The apical portion of the columnar cells of the sternal gland of the termite Zootermopsis nevadensis (Hagen) has been examined with the electron microscope. The cell surface abutting the cuticle is thrown into ridges upon which stand microvilli. Sections show a network of smooth membrane-bound cisternae penetrating the interior of the microvilli. At the bottom of the crevasses between the ridges, an inpocketing of the cell membrane is often found. This is surrounded by a 40-mµ electron-opaque zone that is the insertion of a 22-mµ microtubular component of the cell cytoplasm. The pouch-like structures and their associated microtubules are considered to represent a new cell organelle.
FINE STRUCTURAL CHANGES IN RESPONSE TO HORMONAL STIMULATION OF THE PERFUSED CANINE PANCREAS
Authors: Ichikawa Atsushi
Published: May 2004

The dog pancreas isolated in situ was perfused with oxygenated dog blood and stimulated with pancreozymin, secretin, or both. There were no significant changes in the fine structure of the acinar, centroacinar, or duct cells attributable to the perfusion. Combined glutaraldehyde and osmium fixation gave good preservation of the secretory products of the acinar cell. Before stimulation, the lumen of the acini is filled with material similar in texture to the content of the zymogen granules, but of somewhat lower density. Release of secretion commonly takes place by coalescence of the limiting membrane of zymogen granules with the plasmalemma, but one granule opening at the surface may frequently be joined by others coalescing with its membrane and forming an interconnected series all with contents having the same texture as the released zymogen. Such a mechanism seems to permit a more rapid release of secretory product than discharge of individual granules. Pancreozymin stimulation caused marked depletion of zymogen granules, but no obvious changes in the Golgi apparatus. It is clear, therefore, that this hormone exerts its effect upon release of granules rather than upon their formation. Secretin stimulation of water and bicarbonate secretion caused a marked washing out of the luminal contents, but had little detectable effect on cellular structure.
NUCLEAR MEMBRANES OF CULTURED MAMMALIAN CELLS IN THE PERIOD PRECEDING DNA SYNTHESIS
Authors: Kishimoto Susumu , Lieberman Irving
Published: May 2004

When kidney cells are cultured directly from the rabbit, the nuclear membranes undergo a change that can be measured as an increase in electrophoretic mobility. The change appears to begin immediately upon culture and is maximal 2 hours later, after which the mobility remains constant at the elevated level. Actinomycin D and p-fluorophenylalanine, but not EDTA or ionizing radiation, suppress the increase in nuclear electrophoretic mobility. With synchronously growing L cells, no change was detected in nuclei from cells taken during various parts of the division cycle.
A STUDY OF EXTRACELLULAR SPACE IN CENTRAL NERVOUS TISSUE BY FREEZE-SUBSTITUTION
Authors: Van Harreveld A. , Crowell Jane , Malhotra S. K.
Published: May 2004

It was attempted to preserve the water distribution in central nervous tissue by rapid freezing followed by substitution fixation at low temperature. The vermis of the cerebellum of white mice was frozen by bringing it into contact with a polished silver mirror maintained at a temperature of about -207°C. The tissue was subjected to substitution fixation in acetone containing 2 per cent OsO4 at -85°C for 2 days, and then prepared for electron microscopy by embedding in Maraglas, sectioning, and staining with lead citrate or uranyl acetate and lead. Cerebellum frozen within 30 seconds of circulatory arrest was compared with cerebellum frozen after 8 minutes' asphyxiation. From impedance measurements under these conditions, it could be expected that in the former tissue the electrolyte and water distribution is similar to that in the normal, oxygenated cerebellum, whereas in the asphyxiated tissue a transport of water and electrolytes into the intracellular compartment has taken place. Electron micrographs of tissue frozen shortly after circulatory arrest revealed the presence of an appreciable extracellular space between the axons of granular layer cells. Between glia, dendrites, and presynaptic endings the usual narrow clefts and even tight junctions were found. Also the synaptic cleft was of the usual width (250 to 300 A). In asphyxiated tissue, the extracellular space between the axons is either completely obliterated (tight junctions) or reduced to narrow clefts between apposing cell surfaces.
THE ACROSOME REACTION IN MYTILUS EDULIS
Authors: Niijima Lumiko , Dan Jean
Published: May 2004

The intact acrosome of the Mytilus edulis spermatozoon consists of a conical vesicle, the basal side of which is deeply invaginated so that the whole vesicle forms a sheath around a very slender axial rod, about 2.7 µ long, inserted in a tube passing through the nucleus. The annular base of the acrosomal vesical is filled with a homogeneous substance; the outer wall of the vesicle is lined with a somewhat irregular layer of a particulate substance interspersed with very fine tubular elements, and its lumen is nearly filled by a strand of material which extends from the inner tip of the invagination to the apex of the acrosome. The lumen of the invagination appears empty except for the rod and a delicate sleeve-like structure which surrounds it. The plasma membrane of the sperm cell lies in immediate contact with the acrosomal membrane over its whole outer surface. In its general organization, this molluscan acrosome shows a rather close homology with that of the annelid Hydroides.