Behavior of protoplasm for survival in injured cells ofValonia ventricosa: involvement of turgor pressure

Summary By injuring cells ofValonia ventricosa, one of two survival strategies — wound-healing and protoplast formation — is induced. The present study revealed that turgor pressure, as well as Ca²⁺ in bathing medium, is involved in the choice between these survival strategies. On the process of wound-healing, turgor pressure is recovered in the presence of both the wound plug, which closes the wound immediately after an injury, and the aggregation of protoplasm around the wound, which serves to protect the inflow of outer medium into the protoplasm layer and also to strengthen the wound plug. When the size of the wound is more than 150 m in diameter, the protoplasmic aggregate strengthen the wound plug incompletely and, as a result, wound-healing is unsuccessful. In this case, the ejection of vacuolar sap is repeated, due to partial restoration of turgor pressure. In each ejection, the wound plug is blown off, together with the aggregated protoplasm and, after several ejections are repeated, the cell is unable to heal the wound because of a lack of protoplasm around the wound. Continuous depression of turgor pressure, during the repeat of the unsuccessful wound-healing, induces disorganization of the protoplasm layer. Under these conditions, the centrifugal propagation of protoplasmic aggregation, followed by the protoplasts formation, takes place easily. Effects of turgor pressure and Ca²⁺ in the bathing medium upon the wound healing is discussed and the cytoplasmic behavior for survival of wounded cells is presented schematically.     

VACUOLAR MASS-FLOW IN THE FLORAL HAIRS OF STAPELIA

The flow of vacuolar material was observed in the single-celled hairs of the corolla of Stapelia grandiflora when the tip region of a hair was cut away and the basal region was supplied with water. The vacuolar flow was a bulk flow independent of the concomitant protoplasmic streaming. The observation has been interpreted in terms of Münich's model for the translation of diffusion into mass-flow.     

Latinos in New York: Communities in Transition

Latinos in New York: Communities in Transition. Gabriel Haslip-Viera and Sherrie L. Baver. eds. Notre Dame, IN: University of Notre Dame Press, 1996. 338 pp.     

Llamas, Weavings, and Organic Chocolate: Multicultural Grassroots Development in the Andes and Amazon of Bolivia

"Llamas, Weavings, and Organic Chocolate: Multicultural Grassroots Development in the Andes and Amazon of Bolivia. Kevin Healy. Notre Dame, IN: University of Notre Dame Press, 2001. 482 pp.     

Latest Pleistocene—Holocene paleoceanographic trends on the continental margin of eastern Canada: Foraminiferal,dinoflagellate and pollen evidence

Micropaleontological studies were made of cores from four shelf basins on the eastern Canadian Margin: Emerald and Canso basins on the Scotian Shelf (44°–46° N), Notre Dame Channel, Newfoundland Shelf (50° N) and Cartwright Saddle, Labrador Shelf (55°). Events were correlated using a combination of¹⁴C dates and pollen stratigraphies. Surface- and bottom-water changes were compared on the basis of dinoflagellates and benthic foraminifera, respectively. The results indicate significant paleoceanographic shifts along a north—south gradient both prior to and during the Holocene.Distinct Late Pleistocene—Holocene paleoceanographic events were distinguished in the Emerald, Canso and Notre Dame basins; these events are less obvious in Cartwright Saddle which is in deeper water and further off-shore. Pleistocene glaciomarine sediments in all basins contain a fauna dominated byElphidium excavatum f.clavata; dinoflagellates and pollen are rare or absent. The widespreadElphidium fauna probably reflects turbid glacial meltwater and/or a permanent ice shelf cover from 20,000-10,000 yrs BP. The Notre Dame core also penetrates older sediment with an outer Labrador Current fauna which may represent a late Wisconsinian interstade at about 23,000 yrs BP. From 7,000–10,000 yrs BP a cold water fauna occurred which is similar to modern outer Labrador Current faunas. From about 5000–7000 yrs BP, a warm interval is indicated by a relatively warm-water calcareous benthonic foraminiferal fauna and increased representation of typical Gulf Stream dinoflagellates. The most recent change occurred in the last 2000 years with an abrupt cooling associated with stronger flow of the arctic inner Labrador Current. This cooling event is marked by an increase in arctic dinoflagellates and by an exclusively agglutinated benthonic foraminiferal fauna at two sites (Canso and Notre Dame). These Holocene paleoceanographic changes are not clearly seen in the benthic fauna of the deep northern basin (Cartwright Saddle) although dinoflagellate data at this site indicate that surface-water changes have occurred that are similar to those found in shallower basins.Shifts in the zonal position of the Gulf Stream and changes in the relative mass transports of the West Greenland and Labrador currents are mechanisms which may account for the paleoceanographic events. The glacial—interstadial—glacial sequence recorded in the Notre Dame Channel, in conjunction with other theories on glacial triggering mechanisms, provides biostratigraphic evidence which suggests the onset of a glacial stage in the near future.     

DYNAMICS OF ARBUSCULE DEVELOPMENT AND DEGENERATION IN A ZEA MAYS MYCORRHIZA

A quantitative light and electron microscope study of developing and degenerating mycorrhizal arbuscules of Glomus fasciculatum in Zea mays was carried out in order to estimate three parameters during the colonization cycle. These were: 1) V_v(f,c), the fraction of the host cell volume occupied by a volume of fungus; 2) V_v(cy,c), the fraction of the host cell volume occupied by host cytoplasm; 3) S_v(pr,c), the surface-area-to-volume ratio of the host protoplast to the whole host cell. Uninfected cortical cells had an S_v(pr,c) of 0.13 μm²/μm³. As the fungus penetrates the cell wall, the protoplast invaginates, causing a decrease in protoplast volume and an increase in protoplast S_v. The S_v(pr,c) of a cell containing a mature arbuscule is 1.275 μm²/μm³. Because of the shrinkage of the protoplast, the S_v of the protoplast to its own volume rather than the original cell volume is 2.55 μm²/μm³, or almost a 20-fold increase. Total cell size is unaffected. When the arbuscule is mature, the fungus occupies 42% of the cell, with 24% as 1-μm-diam branches, and 18% as trunk. Arbuscular branch formation progresses at a linear rate and is the most important factor in causing the increased host S_v. The correlation coefficient for V_v(br,c) the volume fraction for arbuscular branches, vs. Sv(pr,c) is r = 0.932 (P < 0.001). Degeneration of the arbuscule is marked by a rapid decrease in branches, host S_v, and host cytoplasm. The trunk develops and degenerates at a slower rate than the branches.     

The relationship of echinate inclusions and coated vesicles on wound healing inNitella flexilis (Characeae)

Summary Wound healing in internodal cells of the freshwater algaNitella flexilis (Characeae) was studied in the light and electron microscope. Immediately after punctation of the cell wall a wound plug is formed which stops outflow of cytoplasm. The plug consists of echinate inclusions which are normally located in the central vacuole. A wound wall consisting of pectin and cellulose microfibrils is formed beneath the plug within one to several hours. During that time the wound shows intensive fluorescence when treated with chlorotetracycline indicating transmembrane Ca²⁺ fluxes. Numerous coated pits and vesicles are found at the plasmalemma. The glycosomes undergo pronounced structural changes. Neither plug nor wound wall formation depend on actin filaments or microtubules as shown by inhibitor experiments with cytochalasin and amiprophos-methyl. The function of the coated vesicles and their interrelationship with other cell organelles is discussed.     

Photobiomodulation alters matrix protein activity in stressed fibroblast cells in vitro

A balance is maintained between matrix synthesis and degradation, and a prolonged increase in matrix metalloproteinases (MMPs) affects healing. Photobiomodulation (PBM) speeds up healing and alters wound environment. The study aimed to determine changes in protein and gene expression of collagen type 1 (Col‐I), MMP‐3 and ‐9 and TIMP‐1 in fibroblasts irradiated at 660 or 830 nm. Commercially purchased human skin fibroblast cells were modeled into five groups namely, normal, normal wounded, diabetic wounded, hypoxic wounded and diabetic hypoxic wounded. Control cells were sham irradiated. Laser irradiation was conducted at 660 or 830 nm (108/or 94 mW, 9.1 cm², 420/or 483 s) with 5 J/cm². Forty‐eight hours post‐irradiation, protein expression of TIMP‐1, MMP‐3, ?9 and Col‐I was determined by flow cytometry and immunofluorescence, and gene expression by real‐time RT‐PCR. There was an increase in TIMP‐1 and Col‐I, and a decrease in MMP‐3 and ‐9, as well as an alteration in mRNA expression of MMP3, MMP9, TIMP1 and COL1A1 in irradiated cells. Due to the responsiveness of the diabetic hypoxic wounded model, the findings propose this model as appropriate for wound healing studies and suggest that PBM promotes the remodeling phase of wound healing by decreasing matrix degradation and upregulating synthesis.      

FROM PROTOPLASM TO SWARMER: REGENERATION OF PROTOPLASTS FROM DISINTEGRATED CELLS OF THE MULTICELLULAR MARINE GREEN ALGA MICRODICTYON UMBILICATUM (CHLOROPHYTA)1

Protoplast regeneration from extruded cytoplasm of the multicellular marine green alga Microdictyon umbilicatum (Velley) Zanardini (Cladophorales, Anadyomenaceae) was investigated. The early process of protoplast formation is comprised of two steps: agglutination of cell organelles into protoplasmic masses followed by generation of a temporary enclosing envelope around them. Agglutination of cell organelles was mediated by a lectin–carbohydrate complementary system. Three sugars, D‐galactosamine, D‐glucosamine, and α‐D‐mannose, inhibited the agglutination process, and three complementary lectins for the above sugars, peanut agglutinin, Ricinus communis agglutinin, and concanavalin A, bound to the surfaces of chloroplasts. Agglutination assay using human erythrocytes showed the presence of lectins specific for the above sugars in the algal vacuolar sap. A fluorescent probe 1‐(4‐trimethylammoniumphenyl)‐6‐phenyl‐a, 3,5‐hexatriene revealed that the envelope initially surrounding protoplasts was not a lipid‐based cell membrane. However, this developed several hours later. Simultaneous fluorescein diacetate and propidium iodide staining showed that the primary envelope had some characteristics of cell membranes, such as semipermeability and selective transport of materials. Also, fluorescein diacetate staining showed esterase activity in the protoplast and relocation of cell organelles and compartmentalization of cytoplasm during the process of regeneration. Both pH 7–9 and salinity 400–500 mM were found to be essentially important for the development of the protoplast envelope. When the basic regeneration process was accomplished, two alternative pathways of development were seen; about 70% of one‐celled protoplasts transformed into reproductive cells within 2 weeks after wounding, whereas others began cell division and grew into typical Microdictyon thalli. Quadriflagellate swarmers were liberated from the reproductive cells, and they germinated into mature individuals. It is therefore suggested that this species may use the wound response as a method of propagation and dispersal.     

The Liberationist Catholic Church in Brazil

Promised Land: Base Christian Communities and the Struggle for the Amazon. Madeleine Cousineau Adriance. Albany: State University of New York Press, 1995. 202 pp.
Women, Religion, and Social Change in Brazil's Popular Church. Carol Ann Drogus. Notre Dame, IN: University of Notre Dame Press, 1997. 226 pp.
Claiming the Virgin: The Broken Promise of Liberation Theology in Brazil. Robin Nagle. New York: Routledge, 1997. 224 pp.
The Brazilian Popular Church and the Crisis of Modernity. Manuel A. Vasquez. Cambridge: Cambridge University Press, 1998. 302 pp.     

The wound response in fresh-cut lettuce involves programmed cell death events

In this work, the involvement of programmed cell death (PCD) in the wound-induced postharvest browning disorder and senescence in butterhead lettuce (Lactuca sativa L.) fresh-cuts was studied. At the wounded (cut, bruised) sites, rapid browning, loss of chlorophyll and massive cell death, accompanied with accumulation of reactive oxygen species and increased electrolyte leakage occurred in a narrow strip of tissue adjacent the injury. The dead cell morphology (protoplast and nuclei shrinkage) together with the biochemical and physiological changes resembled necrotic PCD type. With a slight delay post-wounding, senescence associated with similar cell death features was initiated in distant non-wounded sites. In addition to necrotic PCD, both in wounded and senescing tissue, the appearance of empty cell corpses was observed, indicating that part of the cells might undergo vacuolar PCD (self-digestion of cellular content after vacuole collapse). The wounding-induced local cell death at the primary site of damage suggested that PCD may serve as a mechanism to seal-off the wound by building a physical barrier of dead cells. However, the cell death at sites remote from the wound suggests the distribution of long-distance senescence-inducing wound messengers. Trichomes in unwounded tissue often were the first to show H₂O₂ accumulation and dead cells; thereafter, the elevated H₂O₂ and cell death appeared in connecting cells and senescence progressed over larger areas. This suggests that trichomes may contribute to mediating the wound signalling leading to subsequent senescence. Our findings demonstrate that PCD is an integral part of the wound syndrome in fresh-cut lettuce.     

Wounds incurred in routine cell culture prolong the duration of the life cycle ofAcetabularia acetabulum and require K+ to heal

Summary We have examined the pressure-wound healing response inAcetabularia acetabulum (L.) Silva (Chlorophyta). Commonly incurred in routine cell culture, these wounds induce disruption of the vacuole and translocation of the cytoplasm away from the wound site. Daily wounding of individual cells retarded cytoplasmic healing over time, but had no effect on the rate of membrane healing. The position of the wound along the cell stalk also affected the ability of the cell to heal: cells wounded near the rhizoid healed at least 1.9 times more slowly and were only half as likely to achieve reproduction as were cells wounded either near the apex or at mid-stalk. The 50% mortality of cells wounded at the rhizoid suggests the existence of a physical structure near the primary nucleus which is important to cell viability. The impact of wounding on reproductive potential and time to heal differed with the phase of cell development: juvenile and early adult cells healed 2–2.5 times more quickly but were less likely to achieve reproduction than late adult or reproductive cells. Growth at very high population densities (2.5 cells/ml) impaired the ability of the cells to heal. Growth of cells in seawater containing a range of potassium concentrations revealed that healing depends on potassium and is optimal at a concentration of — 1.51ogM potassium.Abbreviations SE standard error of the mean - LD light/dark - NEM N-ethylmaleimide     

Characterization of clonal populations of theHeliothis zea cell line IPLB-HZ 1075

Summary A total of 24 clones (HZ 1075/UND-A through X) were initially isolated by dilution plating from the established IPLB-HZ 1075 cell line. Many of the isolates with highly vacuolated cytoplasms eventually died during subculturing. The cloned cell strains differed in their predominant morphology, cell doubling times, and relative ability to support replication of the singly encapsulated nuclear polyhedrosis virus ofHeliothis zea (HzSNPV). The origin of the cloned cell strains was confirmed by comparing their isozyme profiles with those of the parental IPLB-HZ 1075 cell line andH. zea larvae using stains for fumerate hydratase, lactate dehydrogenase (LDH), and malic dehydrogenase (MDH). One dipteran and several lepidopteran cell lines maintained in our lab were also separable using stains for LDH and MDH. This research was supported in part by USDA grant number GAM 8400211, and by a Jesse Jones Faculty Research Award from the University of Notre Dame to M. J. F.     

Maintenance of turgor by rapid sealing of puncture wounds in leaf epidermal cells

When leaf epidermal cells are puncture wounded with a glass microcapillary tip, a small droplet of fluid is discharged and then evaporates, leaving a solid residue on the cell surface. For puncture wounds of about 3.5 micrometers in diameter, this process is complete within 2 to 3 seconds. A second puncture wound also exhibits a similar discharge, indicating the persistence of some turgor pressure within the cell, despite damage to the cell wall. Direct measurement of turgor on the large epidermal cells of Tradescantia virginiana L. demonstrated that turgor was substantially maintained (91-96%) after puncture wounding. Anatomical and histochemical evidence suggests that the damaged portion of the cell wall was sealed with an amorphous plug of material comprised of pectinaceous polysaccharides. Rapid sealing of puncture wounds and the maintenance of turgor in epidermal cells may be an important functional component of plant adaptation to physical damage such as that caused by insect feeding.     

Actomyosin is involved in the plasmolytic cycle: gliding movement of the deplasmolyzing protoplast

Summary.  The leaf cells of Chlorophytum comosum seem to have the ability to regulate their protoplast volume and shape during the plasmolytic cycle. This phenomenon was morphologically expressed by the stabilization of the plasmolyzed protoplast volume and shape within 1–5 min after the immersion of the leaf segments in the plasmolytic fluid and temporarily at the onset of deplasmolysis. During the latter stage the plasmolyzed protoplast rounded up and assumed a perfectly convex shape and glided into the cell lumen along the cell axis. This gliding movement was active, nonsaltatory, and conducted with a constant velocity and lasted for a short time. During this movement the protoplast volume did not change appreciably. As far as we know, this movement has not been described so far. Deplasmolysis proceeded and was rapidly completed when the protoplast stopped moving. Leaf cells which have been affected by an antiactin filament drug or myosin inhibitors lost their ability to regulate the volume and shape of the plasmolyzing protoplast. In addition, the gliding protoplast movement was also inhibited in the treated cells. These data show for the first time that the actomyosin system is involved in the mechanism of volume regulation during the plasmolytic cycle and that it underlies the gliding movement of the deplasmolyzing protoplast. Received June 3, 2002; accepted September 26, 2002; published online April 2, 2003 RID="*" ID="*" Correspondence and reprints: Department of Botany, Faculty of Biology, University of Athens, Athens 15784, Greece. E-mail: bgalatis@biol.uoa.gr     

Adherence of Agrobacterium tumefaciens to the cells of immature wheat embryos

Agrobacterium attached to wheat embryos in vitro. This attachment was plasmid independent, and occurred on both wounded and unwounded cell surfaces. The pattern of attachment clearly demonstrated that bacterial attachment to cereal cells follows the same trends observed for dicotyledonous plants. During the inoculation period the bacterial cells attach to the plant cell walls either with lateral or polar orientation. Wounding (mechanical or enzymatic) preferentially promoted adherence of the bacteria at the wound site, however, attachment was not wound dependent.     

STUDIES ON SARGASSUM. I. A LIGHT MICROSCOPIC EXAMINATION OF THE WOUND REGENERATION PROCESS IN MATURE STIPES OF S. FILIPENDULA

Regrowth from wounded stipe explants of Sargassum can be divided into four stages based on cytological changes. The first stage involves changes associated with the wound reactions and the formation of a wound epidermis. The second stage includes the formation of a well defined medullary pit with meristematically active cells around its periphery. Several “bud primordia” are also formed which begin to grow by cell division towards the wound surface. The third stage involves a period of internal tissue differentiation in the “bud primordia” such that mitotic activity is localized in the bud tip and the basal cells grow by cell elongation. The fourth stage marks a major change in the morphology of the regeneration branch from a tubular structure to that of a flattened blade. This change in morphology is preceded by the formation of an apical pit around which the flattened growth appears to be organized.     

Nitrate reductase and respiratory adaptation in Bacillus stearothermophilus

Downey, R. J. (University of Notre Dame, Notre Dame, Ind.). Nitrate reductase and respiratory adaptation in Bacillus stearothermophilus. J. Bacteriol. 91:634-641. 1966.-Bacillus stearothermophilus 2184 required nitrate to grow in the absence of oxygen. Like many facultative microorganisms, the growth obtained anaerobically was considerably less than that obtained aerobically, even though the dissimilatory reduction of nitrate is, in effect, anaerobic respiration. The ability to reduce nitrate depended on the induction of nitrate reductase. Although oxygen at low levels did not retard induction of the enzyme, enzyme synthesis was considerably lessened by aeration. A semisynthetic medium containing nitrate supported aerobic growth of the thermophile but did not support anaerobic growth. The adaptation to nitrate resulted in a decrease in the level of cytochrome oxidase normally present in aerobically grown cells. Although the aerobic oxidation of succinate by the respiratory enzymes from aerobically grown cells was inhibited by 2-N-heptyl-4-hydroxyquinoline-N-oxide, the anaerobic oxidation of succinate by nitrate in a similar preparation from nitrate-adapted cells was not. The nitrate reductase in the bacillus was strongly inhibited by cyanide and azide but not by carbon monoxide. The nitrate reductase catalyzed the anaerobic oxidation of reduced nicotinamide adenine dinucleotide, and appeared to transfer electrons from cytochrome b(1) to nitrate. Cytochrome c(1) did not appear to be involved in the transfer.     

ANALYSIS OF GAMETIC PROTOPLAST RELEASE IN THE CLOSTERIUM PERACEROSUM-STRIGOSUM-LITTORALE COMPLEX (CHLOROPHYTA)1

A biologically active glycoprotein (protoplast-release-inducing protein; PR-IP), which induces the release of gametic protoplasts from mating type minus (mt^-) cells of the Closterium peracerosum-strigosum-littorale complex, was prepared from a medium in which mt^- and mt⁺ cells had been previously incubated together. The process of PR-IP-inducing protoplast release was analyzed. Induction of protoplast release was dependent upon the duration of both PR-IP treatment and preincubation in nitrogen-deficient mating medium before PR-IP treatment. Low cell density in the preculture stage had a significant stimulative effect upon the induction of protoplast release. Light was necessary for protoplast release, especially just before PR-IP treatment. Chloramphenicol and 3-(4-chlorophenyl)-1,1-dimethylurea (CMU) exerted inhibitory effects on protoplast release, especially when they were applied to the preculture stage but not when they were applied to the protoplast-releasing stage after the PR-IP treatment. We suggest that preculture at a low cell density under continuous light conditions that may cause metabolic changes in the chloroplast is a very important stage for gametic protoplast release in this Closterium.     

REGENERATION OF THE SUNFLOWER CAPITULUM AFTER CYLINDRICAL WOUNDING OF THE RECEPTACLE

Using the young capitulum of Helianthus annuus L., a cylindrical plug of undifferentiated receptacle tissue, 1 mm in diameter, was isolated from lateral communication with the rest of the receptacle surface by a vertical circular wound cut, while retaining continuity with the subapical meristem. Within 24 hr, active cell division was induced at the inner and outer surfaces of the wound and in the receptacle epidermis bordering the wound edges, creating a rounded rim at the top of the wound. Within 3–6 days, floral initials, spaced 133–166 μm apart appeared on the flanks of both rims and later on the top of the plug and surrounding receptacle surface. The first formed initials developed into involucral bracts or ray florets and the later ones into disc florets which were organized into contact parastichies, the number of which did not conform with the Fibonacci series. The base of the plug developed into a stem-like structure completing the regeneration of a fully formed functional capitulum. This operation was demonstrated for two sunflower cultivars and occurred in both long and short daylengths.