Tissue Respiration and Mitochondrial Oxidative Phosphorylation of NaCl-Treated Pea Seedlings

The effect of sodium chloride added to root medium of pea seedlings on respiratory activity of tissue segments and on isolated mitochondria was studied. Salinization enhances the respiration of leaves about one-third on a fresh weight, dry weight or protein basis. Roots and stems show only 10 to 15% respiratory stimulation. The onset of respiratory increase in leaves roughly parallels the increase in NaCl content and the decrease in growth rate. At a later stage the elevated respiration is apparent in treated plants even though the concentration of NaCl reaches a plateau and osmotic adjustment is being reached. Stimulation of respiration was found in both etiolated and green plants. Experiments with DNP show that simple uncoupling by salt is not involved; the respiratory increase in control and treated tissue is proportionally the same.

In accordance with increased respiration rates observed in vivo, mitochondria from salt-treated plants show higher rates of oxygen uptake on several substrates. The effect of NaCl added during growth is long term and is distinct from the effect of NaCl added to mitochondria isolated from control plants. Since P/O ratios are not affected by NaCl, the potential for oxidative phosphorylation in salt-affected tissue appears to increase. It is postulated that this increase may lead to changes in ADP and ATP content, and in turn, affect regulation of metabolic pathways.

     

Variations in Lipid and Fatty Acid Content in Relation to Acetyl CoA Carboxylase in the Marine Prymnesiophyte Isochrysis galbana

Lipid and fatty acid content was determined in Isochrysis galbanacultures grown under various environmental conditions in steadystate continuous cultures. Lipid and fatty acid accumulationwas observed under severe nitrogen limiting conditions. Therate of in situ lipid synthesis, determined from ¹⁴C bicarbonateincorporation into lipid fraction, decreased under nitrogenlimited growth (µ<0.72day^–1), concomitant witha reduction in the in vitro activity of the enzyme acetyl CoAcarboxylase. Cellular lipid and fatty acid content remainedfairly constant over a wide range of irradiance levels. Therate of lipid synthesis, however, increased as irra-diance levelwas elevated to a maximal value at a light intensity of 150µmolquanta m^–2s^–1 and slightly decreased at a higherphoton flux. The in vitro activity of ACCase roughly followedthe same pattern of lipid synthesis in response to light intensity.The relative abundance of acetyl CoA carboxylase significantlydecreased in nitrogen limited cultures grown at low dilutionrates (µ<0.72day^–1). A fairly good linear correlationwas measured between the cellular content of ACCase and theenzyme activity in cultures grown under nitrogen limiting conditions.Furthermore, in nitrogen limited cultures, the cellular fattyacid content was linearly related to the cell capacity to producemalonyl CoA, the end product of ACCase and the building blockin fatty acid synthesis. (Received November 20, 1990; Accepted January 17, 1991)     

Thrombin-induced platelet aggregation is affected by external Na+ independently of the Na+/H+ exchange

Thrombin affects blood platelets by activation of Na+/H+ exchange and induction of aggregation, but the relationship between these effects is under debate. The present study attempts to clarify whether the activation of the exchanger activity is required for platelet aggregation. In apparent support of such a requirement, thrombin-induced aggregation is higher in Na+ medium than in N-methylglucamine+ medium and is inhibited by sphingosine, an inhibitor of protein kinase C known to regulate the Na+/H+ exchanger. However, the inhibition of aggregation by sphingosine occurs in both Na+-containing and Na+-free media, the aggregation is identical in Na+ and K+-containing media, and is not inhibited by 5-N-(3-aminophenyl)amiloride, at a concentration 10-fold higher than its Ki for platelet Na+/H+ exchange. Furthermore, at low concentration (0.005 U/ml) thrombin induces aggregation but does not activate the exchange. It is concluded that the activation of Na+/H+ exchange is not required for thrombin-induced platelet aggregation and that the apparent augmentation of aggregation by Na+ is due to an inhibitory effect of N-methylglucamine+.     

Isolation of progenitor cells from cord blood using adhesion matrices

The aim of this study was to develop optimal conditions for selective adhesion and isolation of mesenchymal progenitor cells (MPCs) from cord blood and to determine their potential for osteogenic differentiation. Mononuclear cells (MNCs) were isolated by Ficoll-Paque gradient and plated onto 48-well culture plates precoated with: human or bovine collagen type I, human collagen type IV, fibronectin or matrigel. Cultures were incubated in αMEM containing fetal calf serum. Viability of the adherent cells was determined by alamarBlue® assay after 2, 3, and 4 weeks. After 4 weeks in culture, cells were typsinized and replated. Primary cultures were analyzed by histochemistry and third passage cells by FACS. Isolated fibroblast-like cells were cultured in the presence of osteogenic factors and differentiation determined by Alizarin Red S staining, RT-PCR and electron dispersive spectroscopy (EDS). MNCs adhered to all types of matrices with the greatest adhesion rates on fibronectin. These cells were CD45⁺, CD105⁺, CD14⁺, CD49a⁺, CD49f⁺, CD44⁺ and CD34⁻. The highest incidence of progenitor cells (PC) was observed on fibronectin and polystyrene. Passages were CD45⁻, CD14⁻, CD34⁻ and weakly CD105⁺. Primary cultures expressed endothelial/macrophage RNA markers whether cultured on fibronectin or polystyrene and these markers decreased upon passage. The best osteogenic differentiation was observed in MPCs cultured in osteogenic medium containing vitamin D₃ and FGF9. These cells expressed the bone-related mRNA, collagen type I, core binding factor I (Cbfa I), osteocalcin and osteopontin. EDS of deposits produced by these cells demonstrated a calcium/phosphate ratio parallel to hydroxyapatite. It was concluded that fibronectin increased adhesion rates and isolation potential of cord blood mesenchymal progenitor cells.     

Evolution of fringing reefs: space and time constraints from the Gulf of Aqaba

This study documents the pattern and rate of reef growth during the late Holocene as revealed by unique geological conditions at the subsiding NW Gulf of Aqaba. We discovered that the modern fringing reef near the city Elat grows on top of a fossil submerged mid-Holocene reef platform. Four coral cores from the fossil platform were dated using the radiocarbon and U-Th methods. The fossil corals range from 5.6±0.1 to 2.4±0.03 ka, constraining the initiation of the modern reef to 2,400 years ago at most. We documented the detailed morphology of the reef using aerial photographs and scuba diving. The survey shows that at its northern end, growth of the 2-km-long reef is inhibited by an active alluvial fan, and it is composed of isolated knolls that are just approaching the sea surface. Towards the south, the knolls are progressively larger and closer together, until they form a continuous reef platform. Along this north-to-south trend we follow the evolution of reef morphology, changes in coral distribution, and the development of a lagoon separated from the open sea. Based on these observations, we suggest a four-stage reef growth model: (a) the reef initiates as coral colonies, forms knolls, and begins to grow upward, limited by the sea surface. (b) Upon reaching the surface, the knolls spread laterally, preferentially parallel to the dominant wave direction assuming an elongated morphology. (c) Continued growth results in adjacent knolls eventually coalescing to form a continuous jagged reef. We interpret the spurs-and-grooves morphology that can be traced across the reef at Elat as remnants of the original trends of knolls. (d) While reef expansion continues, the original knoll trends may be obscured as a massive reef front takes shape. Considering reef growth rates and observations from the modern reef at Elat, this evolution scheme predicts an age range of 10³ years for corals on the reef platform. The range and distribution of radiometric ages we obtained from the fossil reef platform underlying the living Elat reef confirm this hypothesis.     

Growth and differentiation of murine cartilage cells in vitro followng a short-term exposure to triamcinolone acetonide

Summary Measurements of ³H-thymidine incorporation, quantitative autoradiography and morphometry were used to evaluate cell behavior during the recovery of mandibular condylar cartilage cultures following short-term exposure to a corticosteroid hormone in vitro. Apical segments of mandibular condyles of newborn mice were initially incubated in the presence of the hormone triamcinolone acetonide (10^-6 M) for 24 h and were thereafter cultured for additional 6 days in hormone-free medium. The present results indicated that the treatment led to a decrease in the rate of incorporation of ³H-thymidine, a feature that lasted for 48 h following the removal of the hormone. Quantitative ³H-thymidine autoradiography of explants that were labeled in the presence of the hormone further substantiated the initial suppressive effect of the hormone on cellular proliferation, a feature that was followed by a recovery. Differences were noted in the pattern of distribution of labeled cells: in control explants, labeled cells progressively moved from the chondroprogenitor compartment into the differentiated portion of the cartilage; in hormone-treated explants, ³H-thymidine labeled cells were confined to the progenitor layer up to 5 days after the treatment and only then appeared in the chondrocytic compartment. The hormone's adverse effect upon differentiation was manifested by both morphology, and by causing a significant increase in the size of the progenitor layer (up to 50.5% on 4th post-treatment day) along with a 70.5% reduction in the size of chondroblastic layer. We conclude that a short-term exposure to a glucocorticoid hormone in vitro interferes with proliferation of chondroprogenitor cells and their subsequent differentiative pathway. While the proliferative activity was restored within 48 h, the hormone's effect on differentiation lasted for a considerably longer period of time.     

Lysine binding to activated human platelets and its similarity to fibrinogen binding

Platelet surface glycoproteins IIb-IIIa are considered to function as the binding site for fibrinogen. Fibrinogen binding is essential for platelet aggregation and several amines have been shown to inhibit this binding. The present study compares the binding properties of 125I-fibrinogen and [3H]lysine with platelets activated by the Ca2+ ionophore A23187. Many lines of similarities in the binding properties are apparent; however, several differences were also found. The similarities are listed below and the differences are pointed out in parentheses. Marked enhancement by platelet activation; deficiency of binding by thrombasthenic platelets lacking the glycoproteins IIb-IIIa; saturability (fibrinogen binding approaches saturation at more than 12 microM, within 10 min; lysine binding at more than 100 mM within 1 min); Ca2+-dependence (at 1 mM Ca2+ lysine binding is minute and fibrinogen binding is half-saturated); reversibility; the binding achieved within 10 min is exchangeable; dissociation depends upon time and external ligand concentration; inhibition by the oligoamines His-Lys and Lys4; inhibition by serum from a thrombasthenic patient who developed anti-glycoproteins IIb-IIIa antibodies; specificity; alanine neither binds to activated platelets nor inhibits fibrinogen binding; it thus appears that the lysine which associates with activated platelets is mostly bound onto the surface of the cells rather than being incorporated. Moreover, the major site of lysine binding seems to be the complexed glycoproteins IIb-IIIa.