bluuebunny/crossref_metadata_2025 huggingface dataset:
ON THE RELATIONSHIP OF LIVER GLUCOSE-6-PHOSPHATASE TO THE PROLIFERATION OF ENDOPLASMIC RETICULUM IN PHENOBARBITAL INDUCTION
Authors: Orrenius Sten , Ericsson Jan L. E.
Published: May 2004

The differentiated effects of phenobarbital treatment on liver microsomal enzymes have been further studied. The relationship between the resulting decrease in the specific glucose-6-phosphatase activity and the enhancement of formation of endoplasmic reticulum membranes with high drug-hydroxylating activity has been investigated with biochemical and histochemical methods. Biochemically and histochemically demonstrable glucose-6-phosphatase activity was found to be present in all endoplasmic reticulum membranes, including the phenobarbital-induced smooth-surfaced proliferates, even though there was an over-all decrease in activity. Actinomycin D did not inhibit the decrease in glucose-6-phosphatase activity. The findings are discussed with reference to the enzyme-membrane relationship in phenobarbital induction.
STUDIES OF SPERMIOGENESIS IN A NEMATODE, NIPPOSTRONGYLUS BRASILIENSIS
Authors: Jamuar Mahendra P.
Published: May 2004

The fine structure of the developing spermatids and the mature sperm of Nippostrongylus brasiliensis was investigated. Immature spermatids are found at one end of the tubelike testis, and the mature sperm at the other. The spermatid has a prominent nucleus, with the chromatin clumped at the margin. It also contains a pair of centrioles, located near the nucleus. The cytoplasm is filled with ribosomal clusters, but it lacks an organized Golgi area or endoplasmic reticulum. Besides the normal mitochondria, the spermatid has specialized mitochondrionlike inclusions with dense matrix, few broad cristae, and a crystalloid structure always facing the nucleus. As spermiogenesis proceeds, the nucleus elongates, comes to lie at one end, and later evaginates to form a separate head structure, leaving the mitochondria and other cytoplasmic organelles in a broad cytoplasmic region. The nuclear material becomes filamentous and spiral, and the centrioles come to lie at one end near the junction of the head and the cytoplasmic portion of the sperm. Microtubules are found in the cytoplasmic region extending from the tubelike nucleus. The specialized mitochondria are about eighteen in number, and are arranged in rows in staggered groups of three around the microtubules in the cytoplasmic region. The mature sperm is aflagellate and lacks an acrosome. No movement of the sperm was ever observed.
OVARIAN STEROID CELLS
Authors: Blanchette E. Joan
Published: May 2004

The granulosa follicle cell of the Graafian follicle of the rabbit ovary differentiates into a lutein cell involved in steroid synthesis. Cytological events which occur within the granulosa cell of the normally stimulated follicle prior to ovulation have been duplicated by the intrafollicular injection of exogenous gonadotrophin. The luteinization of the granulosa cells involves the accumulation of 250- to 300-A, electron-opaque, spherical granules, dispersed within the cytoplasmic matrix, which have been identified as glycogen with the PAS-staining procedure. Further development of the granulosa cell following ovulation involves an increase in cell size, a decrease in the number of RNP particles, and an accumulation of an abundant system of intracellular membranes (agranular endoplasmic reticulum). Glycogen granules first appear in the granulosa cells as the separate, monoparticulate form. After follicle rupture and the formation of agranular endoplasmic reticulum, glycogen particles are present in a rosette arrangement within membrane-bounded vacuoles. The rosette arrangement of glycogen particles is also found dispersed within the cytoplasmic matrix of the lutein cell during the later stages of the cell life-span. Injection of luteinizing hormone or human chorionic gonadotrophin into a mature follicle also produces a marked accumulation of monoparticulate glycogen in the majority of granulosa cells, within 30 min. Cytoplasmic extensions which contain the glycogen masses are noticeably free of RNP particles.
LYSOSOMES IN LYMPHOID TISSUE
Authors: Bowers William E. , Finkenstaedt John T. , de Duve Christian
Published: May 2004

Methods have been developed for the quantitative assay of cytochrome oxidase, esterase, and 11 acid hydrolases in rat-spleen homogenates. These methods seem to be applicable also to other lymphoid tissues. Preliminary studies, extended to nine of the acid hydrolases, indicate that these enzymes occur in partly latent and sedimentable form and that they can be unmasked and rendered soluble by some of the treatments that liberate the enzymes from rat-liver lysosomes. The spleen particles appear to be very sensitive to mechanical injury, a property which necessitates special precautions in homogenizing the tissue. Agglutination of spleen particles takes place to a larger extent in 0.25 M sucrose than in 0.15 M KCl.
STUDIES ON ULTRASTRUCTURAL IDENTIFICATION AND DISTRIBUTION OF PROTEIN-POLYSACCHARIDE IN CARTILAGE MATRIX
Authors: Matukas Victor J. , Panner Bernard J. , Orbison J. Lowell
Published: May 2004

Previous reports on the ultrastructure of cartilage matrix have described fibers, amorphous ground substance and, in some instances, dense matrix granules. The fibers are presumably collagen, but the nature of the granules is unknown. The primary purpose of this study has been to investigate the ultrastructure of cartilage matrix ih chick embryos with particular emphasis on the distribution and composition of these granules. In matrix of the zone of articular cartilage, mature collagen fibers can be seen but granules are not present. In matrix of all other zones of cartilage, fibers are smaller and granules are present. When the matrix of epiphyseal cartilage is compared to that of the zone of hypertrophic cells, fibers are similar but the granules in the latter zone are larger and more numerous. The granules in both zones were digested by hyaluronidase and positive to colloidal iron staining. Chemical analyses of cartilage from these zones indicate the hexosamine and radiosulfate content of the zone of hypertrophic cells to be higher than that of the zone of epiphyseal cartilage. The increased hexosamine was shown by column chromatography to be principally sulfated mucopolysaccharide, thereby indicating a direct correlation between size and number of granules and sulfated mucopolysaccharide content in the two zones. These data and the results of the electron microscopic histochemical studies are consistent with the concept that the granules in cartilage matrix contain acidic mucopolysaccharide.
ISOLATION OF CELL NUCLEI FROM THE MAMMALIAN CEREBRAL CORTEX AND THEIR ASSORTMENT ON A MORPHOLOGICAL BASIS
Authors: Kato Takahiko , Kurokawa Masanori
Published: May 2004

An aqueous method is described for the isolation of highly purified nuclei from the cerebral cortex of adult guinea pigs. Erythrocytes were removed by a short-time perfusion of the brain, myelin fragments by a rapid mechanical method, and blood capillaries by a centrifugal sieving through dense sucrose solutions. The nuclear preparation retained the activity of ATP:NMN adenylyltransferase. Recoveries of DNA in the P4I, P4II, PL and PS preparations were 30, 43, 8, and 7%, respectively. Microscopy and phase contrast microscopy showed a satisfactory removal of erythrocytes, myelin fragments, capillaries, and cytoplasmic elements. Biochemical purity of samples was verified by the absence of several cytoplasmic enzyme activities. In the electron microscope, the majority of nuclei showed well-preserved nuclear membranes, with nuclear pores, and were provided with a finely textured nucleoplasm. Occasional contaminants were elements of endoplasmic reticulum and of the endothelium. Assortment of nuclei on a morphological basis showed that 55–65% and 47–53% of nuclei in the P4I and P4II preparations, respectively, consisted of neuronal nuclei. In the PL preparation, the population of neuronal nuclei ranged between 72 and 83%, while 94–99% of the nuclei in the PS preparation consisted of smaller nuclei, most likely of oligodendroglial origin.
THE SUBMITOCHONDRIAL LOCALIZATION OF MONOAMINE OXIDASE
Authors: Schnaitman Carl , Erwin V. Gene , Greenawalt John W.
Published: May 2004

Controlled osmotic lysis (water-washing) of rat liver mitochondria results in a mixed population of small vesicles derived mainly from the outer mitochondrial membrane and of larger bodies containing a few cristae derived from the inner membrane. These elements have been separated on Ficoll and sucrose gradients. The small vesicles were rich in monoamine oxidase, and the large bodies were rich in cytochrome oxidase. Separation of the inner and outer membranes has also been accomplished by treating mitochondria with digitonin in an isotonic medium and fractionating the treated mitochondria by differential centrifugation. Treatment with low digitonin concentrations released monoamine oxidase activity from low speed mitochondrial pellets, and this release of enzymatic activity was correlated with the loss of the outer membrane as seen in the electron microscope. The low speed mitochondrial pellet contained most of the cytochrome oxidase and malate dehydrogenase activities of the intact mitochondria, while the monoamine oxidase activity could be recovered in the form of small vesicles by high speed centrifugation of the low speed supernatant. The results indicate that monoamine oxidase is found only in the outer mitochondrial membrane and that cytochrome oxidase is found only in the inner membrane. Digitonin treatment released more monoamine oxidase than cytochrome oxidase from sonic particles, thus indicating that digitonin preferentially degrades the outer mitochondrial membrane.
BIOSYNTHESIS OF COLLAGEN
Authors: Fernández-Madrid F.
Published: May 2004

Synthesis of collagen on polyribosomes has been demonstrated in vitro in chick embryo corium by radioisotope incorporation, zone centrifugation through sucrose gradients, and analytical ultracentrifugation. Collagen synthesis was associated with polyribosomes ranging in size, as reflected by their sedimentation constants, from about 180S to approximately 1600S. Most of the newly formed collagen, hydroxyproline, was present on the largest polyribosome aggregates (∼ 350–1600S), but small polyribosomes (∼180–200S) also contained collagen. On the basis of the proline-14C/hydroxyproline-14C ratios and the disrupting effect of collagenase, the proposal is made that the 350–1600S polyribosomes from this tissue are involved predominantly in collagen synthesis. The large polyribosomes are disrupted extensively by collagenase but only partially by ribonuclease and trypsin. Therefore, it appears that they are stabilized by the interaction of newly forming collagen chains. Evidence is presented consistent with the hypothesis that these large polyribosomes are formed by the aggregation of small polyribosomes (180–200S) through the interaction of collagen polypeptides. It is suggested that these small polyribosomes might be involved in the synthesis of subunits of the collagen alpha chain.
AN ARTEFACT IN RADIOAUTOGRAPHY DUE TO BINDING OF FREE AMINO ACIDS TO TISSUES BY FIXATIVES
Authors: Peters Theodore , Ashley Charles A.
Published: May 2004

The binding of labeled free amino acids to liver and to purified protein by commonly used fixatives was investigated. Glutaraldehyde caused 25% of free leucine to be bound to serum albumin in solution, whereas formaldehyde bound only 0.5%. Liver slices were incubated for 2 min in the presence of labeled leucine and of puromycin, which permits absorption of leucine into the cell but inhibits incorporation into protein. Both counting and radioautographic techniques showed that glutaraldehyde bound 30 times, and osmic acid six times, as much free amino acid as did formaldehyde. By comparing liver slices incubated with and without puromycin for 2 min, it was calculated that in radioautographs prepared after fixation with glutaraldehyde, osmic acid, or formaldehyde 63, 25, and 4% respectively of the grains were due to binding of free amino acid. Formaldehyde, freshly prepared from paraformaldehyde, gives good preservation and is the recommended fixative for radioautography. When levels of free substrate in a tissue are high at the time fixative is added, the amount of binding of free substrate induced by the fixative should be included as a control in radioautographic experiments.
HISTOLOGICAL CHANGES DURING THE DEVELOPMENT OF THE BOVINE NUCHAL LIGAMENT
Authors: Wirtschafter Z. T. , Cleary E. G. , Jackson D. S. , Sandberg L. B.
Published: May 2004

The histological changes occurring during the development of the bovine nuchal ligament have been observed in sections of formalin-fixed material from 21 animals ranging in age from 110 days of gestation to 10 yr. The elastic fibers which constitute the bulk of the adult ligament were initially few in number. During fetal development, the fibers showed a rapid increase both in number and in their stainability with the usual elastic stains. The average diameter of these elastic fibers increased only slowly until the last uterine month, at which time it began to increase very rapidly. This rapid rate of increase continued through the first 6 postnatal months, after which the rate of increase slowed markedly. However, the fiber diameter continued to rise steadily throughout the period of the study. During the fetal stage of development, the fibroblastic cells of the ligament exhibited unusual nuclear appearances which distinguish them from other fibroblasts. These consisted of marked clumping of the chromatin and an associated nuclear vacuolation or vesiculation. While these changes seem likely to be artefacts of fixation, their temporal correlation with elastin deposition and their demonstration in other tissue cells engaged in elastin production suggest that the factors responsible for these appearances may be related to elastin synthesis.
DNA REPLICATION PATTERNS AND CHROMOSOMAL PROTEIN SYNTHESIS IN OPOSSUM LYMPHOCYTES IN VITRO
Authors: Schneider Lawrence K. , Rieke William O.
Published: May 2004

DNA replication patterns were determined in the autosomes and sex chromosomes of phytohemagglutinin-stimulated lymphocytes from the opossum (Didelphis virginiana) by employing thymidine-3H labeling and high-resolution radioautography. Opossum chromosomes are desirable experimental material due to their large size, low number (2n = 22), and morphologically distinct sex chromosomes. The autosomes in both sexes began DNA synthesis synchronously and terminated replication asynchronously. One female X chromosome synthesized DNA throughout most of the S phase. Its homologue, however, began replication approximately 3.5 hr later. The two X's terminated DNA synthesis synchronously, slightly later than the autosomes. This form of late replication, in which one X chromosome begins DNA synthesis later than its homologue but completes replication at the same time as its homologue, is apparently unique in the opossum. The male X synthesized DNA throughout S while the Y chromosome exhibited late-replicating characteristics. The two sex chromosomes completed synthesis synchronously, slightly later than the autosomes. Grain counts were performed on all chromosomes to analyze trends in labeling intensity at hourly intervals of S. By analyzing the percent of labeled mitotic figures on radioautographs at various intervals after introduction of arginine-3H, chromosomal protein synthesis was found not to be restricted to any portion of interphase but to increase throughout S and into G2.
PROTEINS IN NUCLEOCYTOPLASMIC INTERACTIONS
Authors: Goldstein Lester , Prescott David M.
Published: May 2004

By the transplantation of amino acid-3H-labeled nuclei between cells and the subsequent isolation of nuclei for quantitative assay, we have confirmed that all the nuclear proteins of Amoeba proteus are divisible into two classes that are sharply defined by their physiological behavior. About 40% of the proteins in the nucleus rapidly migrates back and forth between the nucleus and the cytoplasm. These rapidly migrating proteins (RMP) are 25–50 times more concentrated in the nucleus than in the cytoplasm, and migration into the nucleus therefore occurs against a high concentration differential. The remaining 60% of nuclear proteins has been classified as slow turnover proteins (STP) since (as reported in a following paper) virtually all of them ultimately undergo turnover. Turnover in this context means loss of label from the nucleus, by either protein breakdown or protein migration to the cytoplasm. Isolation of nuclei in the detergent Triton X-100 results in a 20% loss of nuclear proteins but conclusions about RMP and STP were not found to be significantly affected by this loss.
PHAGOCYTOSIS OF LATEX BEADS BY ACANTHAMOEBA
Authors: Korn Edward D. , Weisman Robert A.
Published: May 2004

Electron microscopic studies confirm and extend the conclusions derived previously from a quantitative biochemical study of the phagocytosis of polystyrene and polyvinyltoluene latex beads by Acanthamoeba (1). Latex beads 1.305, 1.90, and 2.68 µ in diameter are ingested individually, with each bead tightly surrounded by a membrane derived from the plasma membrane. Latex beads 0.557, 0.264, 0.126, and 0.088 µ in diameter are accumulated at the surface of the ameba and then phagocytosed, with many beads tightly packed within one membrane-bounded vesicle.
THE TRANSFORMATION OF THE PLASMODIUM GALLINACEUM OOCYST IN AEDES AEGYPTI MOSQUITOES
Authors: Terzakis John A. , Sprinz Helmuth , Ward Ronald A.
Published: May 2004

Sporoblast and sporozoite formation from oocysts of the avian malarial parasite, Plasmodium gallinaceum, after the seventh day of infection in Aedes aegypti mosquitoes offers an interesting example of differentiation involving the appearance and modification of several cellular components. Sporoblast formation is preceded by (a) invaginations of the oocyst capsule into the oocyst cytoplasm, (b) subcapsular vacuolization and cleft formation, (c) the appearance of small tufts of capsule material on the previously noted invaginations, and (d) linear dense areas located just below the oocyst plasma membrane which predetermine the site of emerging sporozoites from the sporoblast. The subcapsular clefts subdivide the once-solid oocyst into sporoblast peninsulae. Within the sporoblast, nuclei migrate from the random distribution seen in the solid oocyst and come to lie at the periphery of the sporoblast just below the linear dense areas noted in the earlier stage. A typical nuclear fiber apparatus occurs in most of the nuclei seen in random sections at this stage although such a fiber apparatus may occasionally be seen in the solid oocyst stage. The nucleus, its associated fiber apparatus, and the overlying dense area appear to induce the onset of sporozoite budding from the sporoblast as well as the formation of the sporozoite pellicular complex and the paired organelle precursor. Several mitochondria are present in each sporozoite, in contrast to the single mitochondrion seen in the merozoites of the erythrocytic and exoerythrocytic stages of avian malaria infection. The paired organelles and associated dense inclusion bodies are formed by condensation of an irregular meshwork of membrane-bound, coarse, dense material. The nature of small, particulate cytoplasmic inclusions is described.
ULTRASTRUCTURE OF THE PRAWN NERVE SHEATHS
Authors: Doggenweiler C. F. , Heuser John E.
Published: May 2004

The sheaths from freshly teased nerve fibers of the prawn exhibit a positive radial birefringence, consistent with their EM appearance as highly organized laminated structures composed of numerous thin cytoplasmic sheets or laminae bordered by unit membranes and arranged concentrically around the axon. The closely apposed membranes in these sheaths are fragile and often break down into rows of vesicles during fixation. Desmosome-like attachment zones occur in many regions of the sheath. The membranes within these zones resist vesiculation and thereby provide a "control" region for relating the type of vesicles formed in the fragile portions of the sheaths to the specific fixation conditions. It is proposed that during fixation the production of artifactual vesicles is governed by an interplay of three factors: (a) direct chemical action of the fixative on the polar strata of adjacent unit membranes, (b) osmotic forces applied to membranes during fixation, and (c) the pre-existing natural relations between adjacent membranes. It is found that permanganate best preserves the continuity of the membranes but will still produce vesicles if the fixative exerts severe osmotic forces. These results support other reports (19) of the importance of comparing tissues fixed by complementary procedures so that systematic artifacts will not be described as characteristic of the natural state.
ISOLATION OF A PROTEIN SUBUNIT FROM MICROTUBULES
Authors: Shelanski M. L. , Taylor E. W.
Published: May 2004

Sea-urchin sperm tails (Strongylocentrotus purpuratus) were obtained by amputation in synthetic sea water and were purified by differential centrifugation. Most of the arms of the outer nine doublets and soluble matrix proteins were removed by this treatment. The central pairs of microtubules were dissolved by dialysis against EDTA at pH 7.5. The extract contained essentially a single component, with a sedimentation constant of 6S, in amounts sufficient to account for the protein content of the central pairs. Incubation of the extract with colchicine-3H gave binding levels approaching 0.5–1.0 mole of colchicine per 105 g protein. Sucrose-gradient analysis showed that the bound-radioactivity profile coincided with the optical-density profile of the 6S protein. It is concluded that the 6S colchicine-binding protein is a subunit of microtubules.
EFFECT OF TEMPERATURE OF ALDEHYDE FIXATION ON THE RADIOAUTOGRAPHIC LOCALIZATION OF RIBONUCLEOPROTEIN IN NUCLEOLI OF HELA CELLS
Authors: Suskind R. Gerald
Published: May 2004

In efforts to clarify the role of the nucleolus and substructures thereof in the assembly or synthesis of protein associated with formation of the complete ribosome, the effect of variation of some conditions of aldehyde fixation on the intranuclear distribution of lysine-3H, arginine-3H, and uridine-3H was studied by differential grain count in radioautographs of PPLO-free HeLa cells. It was found that the nucleolus is a site of rapid assembly or synthesis of a protein, the synthesis of which is inhibited equally by puromycin (200 µg/ml) and by actinomycin D under conditions inhibitory for ribosomal precursor RNA synthesis (P < 0.01). This protein is fixed by phosphate-buffered formalin or glutaraldehyde at pH 7.3, but the label is diminished by fixation in customarily employed acetic ethanol or in formalin at acid pH. Elevation of temperature of formalin or glutaraldehyde fixatives to 37°C consistently reduces the nucleolar protein label, but not the RNA label, by a proportion identical with that incurred by puromycin or actinomycin inhibition. This proportional reduction of nucleolar protein label occurs without evident loss of total grain count and is independent of length of fixation between 30 min and 4 hr, but it is not observed at 23°C. The data support the interpretation that the proportion of nucleolar protein not fixed at 37°C is associated with nucleolar ribosomal RNA but that it is dissociated at 37°C in formalin or glutaraldehyde fixatives, probably on the basis of ionic dissociation of a conjugated ribonucleoprotein.
ACID PHOSPHATASE LOCALIZATION IN RABBIT EOSINOPHILS
Authors: Seeman Philip M. , Palade George E.
Published: May 2004

Eosinophil (and heterophil) leukocytes of glycogen-induced rabbit peritoneal exudates were fixed for 1½ min in 2% glutaraldehyde and examined for acid phosphatase activity both biochemically and cytochemically. Biochemical assays showed that enzymatic activity had been inhibited by only ∼10% under these conditions. The cytochemical reaction in the eosinophil was confined to the granules in which the reaction product appeared in the matrix, not in the crystalline core (or in the core region after the latter's extraction). Granules wherein the matrix was disrupted and the crystalline core degraded or extracted showed the most intense deposition of reaction product, whereas well preserved granules with morphologically intact matrix and crystals were unreactive. Yet, not all disrupted granules gave a positive reaction, indicating that disruption was a necessary but not sufficient condition for reactivity. In many eosinophil leukocytes, most if not all granules were acid phosphatase-positive, provided they had become disrupted to a certain degree. Factors possibly involved in converting the granules from an unreactive to a reactive state are discussed.
ULTRATHIN FROZEN SECTIONS
Authors: Leduc Elizabeth H. , Bernhard W. , Holt S. J. , Tranzer J. P.
Published: May 2004

Endogenous enzyme activity can be readily and routinely demonstrated in ultrathin, frozen sections for electron microscopy. The procedure employed to obtain the best structural preservation as well as enzyme activity in thin sections involved fixation in glutaraldehyde, embedding in thiolated gelatin or pure gelatin, partial dehydration in glycerol, and sectioning in a cryostat at -35°C with a slightly modified Porter-Blum microtome on which the tissue is maintained at -70°C and the knife at -23°C. Kidney cortex was used as test tissue, but a few other organs were occasionally used. Thin sections were floated on the surface of several incubation media routinely employed for enzyme cytochemistry. Positive, specific reactions were obtained for alkaline phosphatase in kidney brush border, for adenosine triphosphatase in brush border and in basal membranes of distal tubules, for acid phosphatase and esterase in lysosomes, and for NADH diaphorase in mitochondria. Mitochondrial ATPase was sporadically evident only in the distal tubule of the kidney. Localizations of enzyme activity reported by other technical approaches were confirmed and in some cases somewhat improved.
THE FORMATION OF THE PRIMARY ENVELOPE DURING OOCYTE DIFFERENTIATION IN TELEOSTS
Authors: Anderson Everett
Published: May 2004

The differentiation of the primary envelope of oocytes of the seahorse (Hippocampus erectus) and the pipefish (Syngnathus fuscus) has been investigated by techniques of light- and electron microscopy. The developing oocytes have been divided into four stages according to size. Oogonia are designated as stage I; stages II and III are oocytes; stage IV represents mature eggs. The primary envelope which is produced by the oocyte is initially a tripartite structure; for convenience of description, the portions are referred to as zones 1, 2, and 3, respectively. Zone 1 first appears as a homogeneous substance at approximately the middle of the long axis of each microvillus. Zone 2 is immediately beneath zone 1 and consists of an extremely electron-opaque granular component. Zone 3 is subjacent to zone 2; it is the largest and most complex of the three. Zone 3 consists of an amorphous material organized in a reticular-like network. Staining procedures indicate that the envelope is composed of a glycoprotein. Just before the oocyte matures there is a structural alteration in zones 2 and 3. Zone 2 becomes a compact, dense layer and zone 3 becomes multilaminate. Subsequent to these changes, zone 1 degenerates. The classification of egg envelopes is discussed, and comparisons are made between the primary envelope of the teleosts investigated and the primary envelopes of other species.
DNA-DEPENDENT FORMATION OF THE SYNAPTINEMAL COMPLEX AT MEIOTIC PROPHASE
Authors: Roth Thomas F. , Ito Michio
Published: May 2004

Evalaution of microsporocytes cultured during discrete periods of meiotic prophase in the presence of deoxyadenosine, an inhibitor of DNA synthesis, indicate that: (1) late leptonema or early zygonema DNA synthesis is required to initiate the formation of the synaptinemal complex; (2) DNA synthesized during late zygonema is necessary for the disjunction of the paired homologs at diplonema; and (3) DNA synthesis in pachynema is a requisite for normal anaphase II separation of sister chromatids.
SYNCHRONOUS MITOCHONDRIAL DIVISION IN NEUROSPORA CRASSA
Authors: Hawley Emerson S. , Wagner R. P.
Published: May 2004

Samples of mycelium of Neurospora crassa of known age were harvested from agar plates and examined with the electron microscope. The relative volume of the mitochondria was determined for mycelium of different ages. The volume measurements indicated that the mitochondria were dividing synchronously in fronts 6, 13, and 22½ hr behind the growing hyphal tips. The sequence of mitochondrial division is hypothesized to include mitochondrial cupping followed by division which results in closely associated daughter mitochondria. On the basis of percentages of mitochondrial cupping and association, mitochondrial division was postulated to be occurring at 6, 14, and 26 hr. Close agreement between the mycelial mass doubling time and the calculated mitochondrial mass doubling time indicates that synchronous mitochondrial division is sufficient to maintain growth. The possibility that mitochondrial division is due to intercellular regulation of a mitochondrial genetic system is advanced.
ANTIGEN-INDUCED CHANGES IN LYMPHOID CELL HISTONES
Authors: Black Maurice M. , Ansley Hudson R.
Published: May 2004

In this study we have examined the solubility of deoxyribonucleoprotein (DNP) isolated from control and antigen-affected thymocytes. 2-M sodium chloride extracts containing the DNP of rat thymus glands were serially diluted. A comparison was made of the effect of dilution on fiber formation in the control and test series. Fiber formation is usually complete for the control material at a salt concentration between 0.63 and 0.57 M. The test material shows some fiber formation within this range. However, a significant portion of the DNP is precipitated at dilutions of 0.54–0.48 M. Ammoniacal silver (A-S) stains the control fibers a characteristic yellowish color. With the test material, those fibers formed within the control range tended to be stained yellowish brown by A-S, whereas those formed only after greater dilution stained blackish. These data, coupled with our previous observations on altered A-S staining, clearly demonstrate an antigen-induced physical and/or chemical alteration of the histone or histone-DNA complex of lymphoid cell chromatin.
ELECTRON MICROSCOPIC EXAMINATION OF SUBCELLULAR FRACTIONS
Authors: Baudhuin Pierre , Berthet Jacques
Published: May 2004

Large granule fractions, containing about 80% of the cytochrome oxidase of the tissue, were isolated from rat liver and used to prepare thin pellicles of packed particles which were submitted to quantitative electron microscopic examination. Various parameters describing the mitochondrial population were determined by measuring the size and number of mitochondrial profiles in sections, and the ratio of the inner to the outer membrane area. The mean particle radius and volume were found to be respectively 0.38 µ and 0.29 µ3; the average areas per mitochondrion were 2 and 5 µ2 for the outer and inner membranes respectively. On the basis of the cytochrome oxidase activity recovered in the particulate fractions, the results were extrapolated to the whole liver, and it was concluded that rat liver contains about 5.1011 mitochondria per gram; this corresponds to a volume of 0.14 ml/g and to an area of 2.5 and 1 m2/g for the inner and outer membranes respectively. The validity and the accuracy of these determinations is discussed and the results are compared to the information which has been obtained by independent methods or by other investigators.
HISTONES OF GENETICALLY ACTIVE AND INACTIVE CHROMATIN
Authors: Comings David E.
Published: May 2004

It has frequently been proposed that a variation in the relative content of lysine-rich, moderately lysine-rich, and arginine-rich histones might provide a mechanism by which specific portions of the genome may be genetically regulated. This possibility was investigated by comparing the electrophoretic pattern of these three fractions in cells differing markedly in their content of genetically active and genetically inactive chromatin. Three models were used: heterochromatin versus euchromatin; metaphase cells versus interphase cells, and mature lymphocytes versus phytohemagglutinin-stimulated lymphocytes. In no case was there a significant difference in the histone patterns of these contrasting models. It is concluded that, although histones may act as a generalized repressor and structural component of chromatin, factors other than a variation in histone pattern may be responsible for repression or derepression of specific segments of the genome.
THE MECHANISM OF MITOCHONDRIAL EXTRUSION FROM PHENYLHYDRAZINE-INDUCED RETICULOCYTES IN THE CIRCULATING BLOOD
Authors: Simpson Charles F. , Kling J. M.
Published: May 2004

The mechanism of mitochondrial extrusion from reticulocytes was studied in whole blood from dogs made anemic by treatment with phenylhydrazine hydrochloride. The initial stage of preparation for mitochondrial extrusion was attraction of vesicles to mitochondria. There was subsequent encirclement of the organelle and other bodies, such as ferritin, by coalesced vesicles forming double membrane-limited vacuoles. Large vacuoles were formed from the union of single vacuoles, and they were usually situated near the periphery of the cell. Fusion of the outer membrane of vacuoles with the plasmalemma of the reticulocyte provided a route for exposure and release of mitochondria and other material to an extracellular location. An extracellular mitochondrion, therefore, was confined by its original double membrane, and a third membrane was derived from the internal boundary of vacuoles.
OBLIQUELY STRIATED MUSCLE
Authors: Rosenbluth Jack
Published: May 2004

Body muscle cells of the bloodworm Glycera, a polychaete annelid, were studied by electron microscopy and compared with muscle cells of the more slowly acting nematode Ascaris, which have been described previously. Both muscles are obliquely striated. The predominant type of bloodworm fiber is characterized by a prominent transversely oriented sarcoplasmic reticulum with numerous dyads at the surface of each cell. Thick myofilaments are ∼3 µ long and overlap along ∼60% of their length in extended fibers and ∼80% in shortened fibers. There is virtually no endomysium and very little intracellular skeleton, and the cells are attached by desmosomes to one another rather than to connective tissue. Dense bodies are absent from the fibers and in their place are Z lines, which are truly linear rather than planar. Scattered among the predominant fibers are others, less orderly in arrangement, in which the SR is much less prominent and in which the thick filaments are thicker and longer and overlap to an even smaller degree. It is suggested that physiological differences between bloodworm and Ascaris muscles derive from differences in the proportion of series to parallel linkages between the contractile elements, differences in the amount and disposition of the SR, and differences in the impedance to shear within the myofibrils.
RIBONUCLEOPROTEIN PARTICLES IN THE AMPHIBIAN OOCYTE NUCLEUS
Authors: Rogers M. Elizabeth
Published: May 2004

Studies of the sedimentation properties of RNP1 material from the nucleus of the amphibian oocyte have indicated (1) that there are few, if any, 78S ribosomes in the nucleus, (2) that there are smaller particles sedimenting at 50-55S and 30S, and (3) that the larger of these is the precursor of the 60S subunit of the cytoplasmic ribosomes. Although the nature of the 30S material is not completely clear, it probably includes precursor particles to the 40S ribosomal subunit. Heavy (50-55S) particles are predominant in immature oocytes of Triturus viridescens, whereas in immature oocytes of Triturus and Amblystoma mexicanum they are reduced greatly in amount, but are still detectable. Double-labeling studies of RNA and protein reveal that both types of particle incorporate uridine-3H, but that the 50-55S material of immature oocytes does not incorporate 14C-labeled amino acids. However, other evidence exists that favors the RNP nature of this material. Sedimentation analyses after SDS extraction show that 50-55S particles contain 40 and 30S RNA, whereas 30S particles contain 20S RNA. These types of RNA represent at least 80% of all the extractable nuclear RNA. The 50-55S particles are probably heterogeneous, including both particles containing mostly 40S RNA and particles containing only 30S RNA.
CALCIUM METABOLISM IN HELA CELLS AND THE EFFECTS OF PARATHYROID HORMONE
Authors: Borle André Bernard
Published: May 2004

Calcium metabolism was investigated in HeLa cells. 90% of the calcium of the cell monolayer is bound to an extracellular cell coat and can be removed by trypsin-EDTA. The calcium concentration of the naked cell, freed from its coat, is 0.47 mM. The calcium concentration of the medium does not affect the concentration of the naked cell calcium. However, the calcium of the cell coat is proportional to the calcium concentration in the medium. Calcium uptake into the cell coat increases with increasing calcium concentration of the medium, whereas uptake by the naked cell is independent of the calcium of the medium. Anaerobic conditions and metabolic inhibitors do not inhibit calcium uptake by the cell, a fact suggesting that this transfer is a passive phenomenon. The calcium in the extracellular cell coat, was not affected by parathyroid hormone. In contrast, the hormone increased the cellular calcium concentration by stimulating calcium uptake or by enhancing calcium binding to some cell components. These results suggest that, contrary to current thinking, parathyroid hormone influences the cellular calcium balance by mobilizing calcium from the extracellular fluids in order to increase its concentration in some cellular compartment. It is proposed that these effects can enhance calcium transport.
LOCALIZATION OF HYALURONIC ACID IN SYNOVIAL CELLS BY RADIOAUTOGRAPHY
Authors: Barland Peter , Smith Carol , Hamerman David
Published: May 2004

Cultured human synovial cells secrete hyaluronic acid (HA) into the culture medium. Glucosamine-6-3H was shown to be a direct and relatively specific precursor of HA-3H by the following observations: the susceptibility of nondialyzable radioactivity in the medium to hyaluronidase, its migration with hexuronic acid on zone electrophoresis in polyvinyl chloride, its exclusion from Sephadex G-200, and the localization of radioactivity to glucosamine after hydrolysis of the labeled polysaccharide. The presence of intracellular HA-3H was established by sequential extraction of labeled cells and by radioautography of synovial cell cultures digested with hyaluronidase in situ. When cells were exposed to medium lacking glucose, glucosamine-3H-uptake was enhanced; and this made possible electron microscopic radioautographic studies. These studies demonstrate the early and continued presence of HA-3H within the Golgi apparatus.
DISTRIBUTION OF PHOSPHATASES IN THE GOLGI REGION AND ASSOCIATED STRUCTURES OF THE THORACIC GANGLIONIC NEURONS IN THE GRASSHOPPER, MELANOPLUS DIFFERENTIALIS
Authors: Lane Nancy J.
Published: May 2004

The neuronal perikarya of the grasshopper contain sudanophilic lipochondria which exhibit an affinity for vital dyes. These lipochondria are membrane-delimited and display acid phosphatase activity; hence they correspond to lysosomes. Unlike those of most vertebrates, these lysosomes also hydrolyze thiamine pyrophosphate and adenosine triphosphate. Like vertebrate lysosomal "dense bodies," they are electron-opaque and contain granular, vesicular, or lamellar material. Along with several types of smaller dense bodies, they are found in close spatial association with the Golgi apparatus. The Golgi complexes are frequently arranged in concentric configurations within which these dense bodies lie. Some of the smaller dense bodies often lie close to or in association with the periphery of dense multivesicular bodies. Further, bodies occur that display gradations in structure between these multivesicular bodies and the dense lysosomes. Acid phosphatase activity is present in the small as well as the larger dense bodies, in the multivesicular bodies, and in some of the Golgi saccules, associated vesicles, and fenestrated membranes; thiamine pyrophosphatase is found in both the dense bodies and parts of the Golgi complex. The close spatial association of these organelles, together with their enzymatic similarities, suggests the existence of a functional or developmental relationship between them.
STUDIES ON LYSOSOMES
Authors: Hirschhorn Rochelle , Brittinger Günter , Hirschhorn Kurt , Weissmann Gerald
Published: May 2004

Subcellular fractions were isolated by differential centrifugation from pure suspensions of human blood lymphocytes incubated with and without phytohemagglutinin (PHA). Between 30 and 120 min after addition of PHA to intact cells, redistribution of acid hydrolases (beta glucuronidase, acid phosphatase), from a 20,000 g x 20 min granular fraction into the corresponding supernatant, was observed. No increase in total acid hydrolase activity was found at these times. The mitochondrial marker enzyme, malate dehydrogenase, did not undergo redistribution. Granules derived from PHA-treated cells became more fragile upon subsequent incubation with membrane-disruptive agents in vitro (streptolysin S, filipin). These changes were associated with an increase in the over-all permeability of the stimulated cell to substances in the surrounding medium, such as neutral red. Augmentation of dye entry into lymphocytes required intact metabolism as judged by response to temperature and inhibitors (cyanide, antimycin A, 2,4-dinitrophenol). PHA, however, did not release enzyme activity from hydrolase-rich granules in vitro or render them more susceptible to subsequent challenge with membrane-disruptive agents. These studies suggest that PHA induces early changes in the surface of lymphocytes. The consequent redistribution of acid hydrolases may play a role in remodeling processes of the stimulated cells.
OOCYTE DIFFERENTIATION IN THE SEA URCHIN, ARBACIA PUNCTULATA, WITH PARTICULAR REFERENCE TO THE ORIGIN OF CORTICAL GRANULES AND THEIR PARTICIPATION IN THE CORTICAL REACTION
Authors: Anderson Everett
Published: May 2004

This paper presents morphological evidence on the origin of cortical granules in the oocytes of Arbacia punctulata and other echinoderms. During oocyte differentiation, those Golgi complexes associated with the production of cortical granules are composed of numerous saccules with companion vesicles. Each element of the Golgi complex contains a rather dense homogeneous substance. The vesicular component of the Golgi complex is thought to be derived from the saccular member by a pinching-off process. The pinched-off vesicles are viewed as containers of the precursor(s) of the cortical granules. In time, they coalesce and form a mature cortical granule whose content is bounded by a unit membrane. Thus, it is asserted that the Golgi complex is involved in both the synthesis and concentration of precursors utilized in the construction of the cortical granule. Immediately after the egg is activated by the sperm the primary envelope becomes detached from the oolemma, thereby forming what we have called the activation calyx (see Discussion). Subsequent to the elaboration of the activation calyx, the contents of cortical granules are released (cortical reaction) into the perivitelline space. The discharge of the constituents of a cortical granule is accomplished by the union of its encompassing unit membrane, in several places, with the oolemma.
EFFECT OF ACRIFLAVIN ON THE KINETOPLAST OF LEISHMANIA TARENTOLAE
Authors: Simpson Larry
Published: May 2004

The loss of kinetoplast DNA in Leishmania tarentolae, which occurs in the presence of low concentrations of acriflavin, was found to be a result of selective inhibition of replication of this DNA. Nuclear DNA synthesis was relatively unaffected and cell and kinetoplast division proceeded normally for several generations. An approximately equal distribution of parental kinetoplast DNA between daughter kinetoplasts resulted in a decrease in the average amount of DNA per kinetoplast. The final disappearance of the stainable kinetoplast DNA occurred at a cell division in which all the remaining visible kinetoplast DNA was retained by one of the daughter cells. The selective inhibition of kinetoplast DNA synthesis was caused by a selective localization of acriflavin in the kinetoplast. The apparent intracellular localization of dye and the extent of uptake at a low dye concentration could be manipulated, respectively, by varying the hemin (or protoporphyrin IX) concentration in the medium and by adding red blood cell extract (or hemoglobin). Hemin and protoporphyrin IX were found to form a complex with acriflavin. During growth in acriflavin, cells exhibited an increasing impairment of colony-forming ability and rate of respiration. No change in the electrophoretic pattern of total cell soluble proteins was apparent. The data fit the working hypothesis that the loss of kinetoplast DNA leads to a respiratory defect which then leads to a decrease in biosynthetic reactions and eventual cell death. A possible use of the selective localization of acriflavin in the kinetoplast to photooxidize selectively the kinetoplast DNA is suggested.
THE DISTRIBUTION OF DNA AMONG DIVIDING MITOCHONDRIA OF TETRAHYMENA PYRIFORMIS
Authors: Parsons John A. , Rustad Ronald C.
Published: May 2004

A squash technique was developed for log phase Tetrahymena pyriformis which permitted the resolution of over 100 individual mitochondria from a single cell. Mitochondria incorporated thymidine at all stages of the cell cycle, even when nuclear DNA synthesis was not occurring. During the stage of macronuclear DNA synthesis, however, there was a significant increase in the extent of mitochondrial labeling. Low radioautograph background suggests that mitochondrial DNA is synthesized at the mitochondria themselves. All mitochondria incorporated thymidine-3H within one population-doubling time. Grain counts also showed that the amount of mitochondrial label was retained for four generations and that this label remained randomly distributed among all mitochondria during this time. The results are not consistent with any theory of de-novo or "microbody" origin of mitochondria, but do support the hypothesis that mitochondria are produced by the growth and division of preexisting mitochondria. The stability of the mitochondrial DNA and its distribution among daughter mitochondria satisfy two prerequisites for a genetic material. The possibility is discussed that some of the genetic information for the mitochondrion is contained in the DNA associated with this organelle.