Characterization of the pyrenoid isolated from unicellular green algaChlamydomonas reinhardtii: Particulate form of RuBisCO protein

Summary The pyrenoid is a protein complex in the chloroplast stroma of eukaryotic algae. After the treatment with mercury chloride, pyrenoids were isolated by sucrose density gradient centrifugation from cell-wall less mutant cells, CW-15, as well as wild type cells, C-9, of unicellular green algaChlamydomonas reinhardtii. Pyrenoids were characterized as a fraction whose protein/chlorophyll ratio was very high, and also examined by Nomarski differential interference microscopy. Most of the components consisted of 55 kDa and 16 kDa polypeptides (11) which were immunologically identified as the large and small subunit of RuBisCO (ribulose-1,5-bisphosphate carboxylase/oxygenase) protein, respectively. Some minor polypeptides were also detected. Substantial amount of RuBisCO protein is present as a particulate form in the pyrenoid in addition to the soluble form in algal chloroplast stroma.Abbreviations BPB bromophenol blue - DAB 3,3-diaminobenzidine - DTT dithiothreitol - ELISA enzyme-linked immunosorbent assay - High-CO₂ cells cells grown under air enriched with 4% CO₂ - Low-CO₂ cells cells grown under ordinary air (containing 0.04% CO₂) - NP-40 nonionic detergent (Nonidet) P-40 - PAGE polyacrylamide gel electrophoresis - PAP peroxidase-antiperoxidase conjugate - RuBisCO ribulose-1,5-bisphosphate carboxylase/oxygenase - RuBP ribulose-1,5-bisphosphate - SDS sodium dodecylsulfate     

Ultrastructural analysis ofErysiphe graminis haustoria and subcuticular stroma ofVenturia inaequalis using cryosubstitution

Summary Cryosubstitution provides an improved ultrastructural preservation of the two plant pathogensVenturia inaequalis andErysiphe graminis when compared to conventional preparation methods. Further, freezing the infected whole leaf material on a copper block cooled with liquid helium gave better results than those observed with the propane plunging method. Novel observations concerning the fungal stroma and haustoria were made which showed ribosomes organized into groups that were evenly distributed throughout the cytoplasm of both fungi. Stretches of rough endoplasmic reticulum were present, and microtubules were seen, sometimes associated with mitochondria. A large number of darkly staining vacuoles were observed in both fungi. The polarity of organelles and microtubules along the longitudinal axis of the haustorial body ofE. gramnis and along the growing direction of subcuticular stroma and runner hypha ofV. inaequalis was evident. InE. graminis filasomes were observed, as were Golgi-like bodies. These new observations, together with the advantages of the cryosubstitution technique, can serve as a basis for further studies in understanding host-parasite interactions.     

Ultrastructure de l'endomètre humain normal

Résumé L'étude du chorion cytogène de 15 endomètres humains normaux prélevés à divers moments du cycle menstruel a précisé d'importantes variations des vaisseaux et des cellules.Les vaisseaux subissent une maturation progressive: en phase proliferative moyenne, des pointes d'accroissement se forment à partir du réseau vasculaire profond activé; au milieu du cycle, elles se transforment en capillaires typiques; en phases sécrétoires moyenne et avancée, des péricytes migrent dans le chorion cytogène et se différencient en cellules choriales.Ces cellules choriales, dites fixes, ont une évolution biphasique au cours du cycle menstruel. Au milieu de chacune des phases, proliférative et surtout sécrétoire, les cellules choriales fixes, dites alors fibroblastoïdes, montrent une intense activité de synthèse. A ces périodes de synthèse succède une involution cellulaire, peu marquée en fin de phase proliferative, intense en phase sécrétoire avancée.Les cellules dites prédéciduales sont des cellules choriales fixes involuées et hyperhydratées; elles vont, en phase menstruelle, évoluer de plusieurs façons: la plupart d'entre elles régénèrent, certaines se nécrosent focalement ou totalement, d'autres font preuve d'activité macrophagique, en particulier collagénolytique.

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Ultrastructure of the normal human endometriumI. The stroma

Summary 15 human endometria during the normal menstrual cycle have been investigated. Important alterations of the vessels and the stroma cells occur.The vessels are the site of gradual maturation. In the mid proliferative phase, growing capillaries rise from the deep-seated vascular system. In the middle of the cycle, they change into typical capillaries. In the mid and late secretory phases, pericytes leave the walls of the capillaries and differentiate into stroma cells.These stroma cells undergo a biphasic cyclic evolution. The middle of the proliferative and particularly of the secretory phase is marked by an intensive synthetic activity of the stroma cells which are called, at this time, fibroblastoïd stroma cells. These two periods of synthesis are followed by cellular involution, mild in the proliferative, intense in the secretory phase.The so-called predecidual cells are hyperhydrated involuted stroma cells. In the menstrual phase they behave very differently of: the majority regenerates, some predecidual cells are the site of focal or total necrosis, others show a macrophagic activity which is conspicuous in some cells having a collagenolytic activity.

Nous remercions Monsieur le Professeur Gandar, Directeur de la Clinique Gynécologique et Obstétricale 1 de la Faculté de Médecine de Strasbourg de nous avoir confié, pour examen ultrastructural, le matériel nécessaire à cette étude.     

Regulation of mammary epithelial cell function: a role for stromal and basement membrane matrices

Summary Interactions between epithelial cells and their environment are critical for normal function. Mammary epithelial cells require hormonal and extracellular matrix (ECM) signalling for the expression of tissue specific characteristics. With regard to ECM, cultured mammary epithelial cells synthesize and secrete milk proteins on stromal collagen I matrices. The onset of function coincides both with morphogenesis of a polarized epithelium and with deposition of basement membrane ECM basal to the cell layer. Mammary specific morphogenesis and biochemical differentiation is induced if mammary cells are cultured directly on exogenous basement membrane (EHS). Thus ECM may effect function by the concerted effect of permissivity for cell shape changes and the direct biochemical signalling of basement membrane molecules.A model is discussed where initial ECM control of mammary epithelial cell function originates in the interstitial matrix of stroma and subsequently transfers to the basement membrane when the epithelial cells have accumulated and deposited an organized basement membrane matrix.Dedicated to Professor Stuart Patton on the occasion of his 70th birthday.     

Control of photosynthesis in leaves as revealed by rapid gas exchange and measurements of the assimilatory force FA

The rapid transients of CO₂ gas exchange have been measured in leaves ofHelianthus annuus L. In parallel experiments the assimilatory force F_A, which is the product of the phosphorylation potential and the redox ratio NADPH/NADP, has been calculated from measured ratios of dihydroxyacetone phosphate to phosphoglycerate in the chloroplast stroma and in leaves. The following results were obtained: (i) When the light-dependent stroma alkalization was measured under steady-state conditions for photosynthesis in air containing 2000 μl · l^-1 CO₂, alkalization increased with photosynthesis as the quantum flux density (irradiance) was increased. This contrasts to the light-dependent stroma alkalisation measured in dark-adapted leaves during the dark-light transient (Laisk et al. 1989, Planta177, 350–358) which reached a maximum at a quantum flux density far below that necessary to saturate photosynthesis. This maximum was about three times higher than the maximum stroma alkalization at light- and CO₂-saturated photosynthesis. (ii) Accurate calculations of the assimilatory force F_A require a consideration of the stromal pH. However, under many conditions, changes in the stromal pH resulting from changes in photosynthetic flux can be neglected because they are small. (iii) Stromal ratios of dihydroxyacetone phosphate to phosphoglycerate are generally lower than ratios measured in leaf extracts. The value of F_A calculated from stromal metabolites was about 30% lower than F_A calculated from cellular metabolites. Still, it appears sufficient for many purposes to calculate F_A from metabolite measurements in leaf extracts. (iv) In the light, the catalytic capacity of the photosynthetic apparatus is adjusted to the level of irradiance. The response of carbon assimilation to large increases in irradiance is slow because it requires enzyme activation. Deactivation of the Calvin cycle induced by decreases in irradiance is slower than activation. (v) Changes in catalytic capacity and in the availability or level of substrates such as CO₂ alter the flux resistance of the Calvin cycle. A decrease in flux resistance explains why F_A often does not increase by much and may actually decrease when carbon flux is increased. Adjustments of flux resistances in the Calvin cycle and of photosystem-II activity in the electron-transport chain permit varying rates of photosynthesis at low levels of ATP and NADPH. As NADP remains available, the danger of over-reduction which leads to photoinactivation of electron transport is minimized. K.R. und U.H. were guests of the Estonian Academy of Sciences. Support by the Estonian Academy of Sciences, the Sonderforschungsbereich 251 of the University of Würzburg and the Fonds der Chemischen Industrie is gratefully acknowledged.     

二次碱化法制备羧甲基茯苓多糖

本研究采用二次碱化新工艺制备了水溶性羧甲基茯苓多糖,并对新旧工艺进行了比较。结果表明,使用改进的新工艺完全可以制备出质量符合要求的产品。     

Studies on bone marrow histogenesis: Morphometric and autoradiographic studies of regenerating marrow stroma in extramedullary autoimplants and after evacuation of marrow cavity

Summary The marrow cavity of the rat tibia was mechanically evacuated and autoimplanted to the subcutaneous tissue. The regenerative process which restored the integrity of marrow stroma and hemopoiesis, was morphometrically evaluated in whole mount of tibia. Following evacuation, the clot filled the cavity. The granulation tissue then appeared and expanded, penetrating and replacing the clot. The fibroblasts of the granulation tissue differentiated into osteoblasts forming osteoid bone. Within its interstices, the primordial marrow consisting of loose connective tissue and vascular sinuses appeared and hemopoiesis resumed. Expansion of hemopoiesis resulted in the resorption of bone and within three weeks the tibial cavity was restored to the pre-evacuation state.Autoradiography indicated that the labeling index was initially high in fibroblasts and osteoblasts but was subsequently reduced while it increased in osteocytes, cells of Haversian canals, stromal and hemopoietic cells of marrow. The finding is in disagreement with the view that the regenerative process originates from the Haversian canal. When the label was introduced on day 4 post-operatively, it subsequently appeared in osteocytes, cells of Haversian canal, stromal elements of the marrow, but not in the hemopoietic cells. This indicates complete dissociation of marrow stroma and hemopoietic stem cell.Supported by NASA Contract NSG 9061. Mehdi Tavassoli is the recipient of a CRD Award AM-70551     

Ecological photodynamic therapy: New trend to disrupt the intricate networks within tumor ecosystem

As with natural ecosystems, species within the tumor microenvironment are connected by pairwise interactions (e.g. mutualism, predation) leading to a strong interdependence of different populations on each other. In this review we have identified the ecological roles played by each non-neoplastic population (macrophages, endothelial cells, fibroblasts) and other abiotic components (oxygen, extracellular matrix) directly involved with neoplastic development. A way to alter an ecosystem is to affect other species within the environment that are supporting the growth and survival of the species of interest, here the tumor cells; thus, some features of ecological systems could be exploited for cancer therapy. We propose a well-known antitumor therapy called photodynamic therapy (PDT) as a novel modulator of ecological interactions. We refer to this as “ecological photodynamic therapy.” The main goal of this new strategy is the improvement of therapeutic efficiency through the disruption of ecological networks with the aim of destroying the tumor ecosystem. It is therefore necessary to identify those interactions from which tumor cells get benefit and those by which it is impaired, and then design multitargeted combined photodynamic regimes in order to orchestrate non-neoplastic populations against their neoplastic counterpart. Thus, conceiving the tumor as an ecological system opens avenues for novel approaches on treatment strategies.