Simultaneous measurements of the intra- and extra-cellular oxygen concentration in viable cells.

An EPR method that can measure the intra- and extra-cellular oxygen concentration [O2] simultaneously in vitro has been developed using specially designed nitroxides. In the presence of Fe(CN)6(3-) in the medium, intracellular [O2] is measured by a neutral 15N-nitroxide and extracellular [O2] is measured by a negatively charged 14N-nitroxide, since charged species do not enter cells and the EPR spectrum of a 15N-nitroxide does not overlap with that of a 14N-nitroxide. The method is based in part on the minimal broadening of negatively charged nitroxides by Fe(CN)6(3-) and the very effective broadening of neutral nitroxides by the same paramagnetic ions. Results with this method confirm the existence of gradients in [O2] between the extracellular and intracellular compartments in CHO cells and M5076 tumor cells, even without stimulation of cellular respiration by CCCP. The nature of the barrier that needs to be involved to account for the experimental results raises some significant questions.     

Mechanobiology of conjunctival epithelial cells exposed to wall shear stresses

Biomechanics and Modeling in Mechanobiology - The human conjunctival epithelial cells (HCEC) line the inner sides of the eyelids and the anterior part of the sclera. They include goblet cells that...     

Top predator fish assemblages in Northern Patagonia,Argentina. What factors regulate their patterns of distribution and abundance?

According to assemblage theory, three factors regulate fish biogeography: restriction of dispersion, environmental restrictions and biotic interactions. The first two factors act on a regional scale and delimit the area of action of the third, which operates on a local scale. Salmonid introductions began in Patagonia in 1904, and this has led to a restructuring of trophic webs and an increase in the number of top predators. This situation allowed us to evaluate, in a natural setting, how communities are formed on different geographic scales. We studied two large basins in Patagonia, situated close to each other but with different assemblages of top predatory fish. We hypothesized that differences in the structuring of the top predator assemblages between and within the basins are due to 1) environmental factors and dispersion processes facilitated by connectivity on a regional scale; and 2) biotic interaction (internal dynamics) between native perch and salmonids, the former acting as a modulator of the top predator assemblages on a local scale. To test these hypotheses, we analysed the top predator assemblages of 16 lakes and one reservoir, as well as their environmental characteristics. We performed a cluster analysis and related the resulting assembly groups to environmental factors by means of a tree model. We also analysed fish diets, using a similarities test to study biotic interactions. On regional and local scales, water basin, degree of connectivity, area, temperature and Zoogeographic integrity coefficient (ZIC) were important factors in the structuring of top predator assemblages. On a local scale, creole perch modulates the salmonid populations through feeding and the consequent distribution of resources. Our work showed that the structure of top predator assemblages was determined by a combination of local and regional factors acting in synergy, as postulated by the assemblage theory.     

In the search for new anticancer drugs XII. Synthesis and biological evaluation of spin labeled nitrosoureas

The spin labeled nitrosourea 1-(2-chloroethyl)-3-(1-oxyl-2,2,6,6- tetramethyl-piperidinyl)-1-nitrosourea (SLCNU, 4) and its analogues 5-7 were synthesized either by a regio-selective method or by a conventional route via the nitrosation of the spin labeled intermediates (11a-e). Nitrosation of the ureas 11a-e with dinitrogen tetraoxide resulted in better yields than those obtained with sodium nitrite. The nitrosoureas 4-8 were tested for their anticancer activity against the lymphocytic leukemia P388 in mice. Thus, either at the equal molar dose or at the dose of equal toxicity level, the SLCNU (4) was found to be more active than the clinically used CCNU (1). Unlike CCNU (1) whose LD50 is 56 mg/kg, the SLCNU (4) possesses a low toxicity (LD50 123 mg/kg). Therefore, SLCNU (4) is a promising new entry into the nitrosourea class of anticancer drugs.     

Development of nitroxides for selective localization inside cells

The use of nitroxides to measure intracellular phenomena, especially oxygen concentrations, is a new and potentially important approach to a number of physiological and pathophysiological studies. This study provides data indicating the feasibility of developing nitroxides that localize selectively in the intracellular compartment; it is based on the use of readily hydrolysed ester linkages, such that the nitroxides become converted intracellularly to ionic derivatives that do not cross cell membranes readily. Up to 120-fold increased concentrations of intracellular nitroxides (and their one electron reduction product, the hydroxylamines) were obtained. The ESR spectra of the intracellular nitroxides were consistent with their conversion to the ionic species. Preliminary studies indicate that these nitroxides have the properties needed for their use as probes of intracellular concentrations of oxygen and that it should be feasible to synthesize nitroxides that will be even more effective for this purpose.     

In the search for new anticancer drugs. X. N,N; N',N'-bis(1,2-ethanediyl)-N'-(1-oxyl-2,2,6,6-tetramethyl- 4-piperidinylaminocarbonyl) phosphoric triamide--a new potential anticancer drug of high activity and low toxicity

A new nitroxyl labeled TEPA derivative 5 containing the urea bridge between the phosphorus and the nitroxyl moiety, and the congeners containing the NOH and NH groups instead of the nitroxyl function were synthesized, and tested in vivo on CD2F1 mice for anticancer activity against P388 and L1210. The nitroxyl compound is more active than the reduced forms. The nitroxyl compound 5 elicits 170% ILS at 90 mg/kg after 30 days and 439% ILSmax after 60 days against P388, and has a higher therapeutic ratio (26.4) than the clinically used Thio-TEPA (2.75). The LD50 of 5 is 270 mg/kg, while that of Thio-TEPA is 18 mg/kg. Consequently, the nitroxyl compound 5 is a promising new anticancer drug.     

Sucking herbivore assemblage composition on greenhouse <Emphasis Type="Italic">Ficus</Emphasis> correlates with host plant leaf architecture

Understanding arthropod herbivore selectivity trends towards host plant attributes is essential for predicting plant-associated herbivore assemblage structure. Little is known about such interactions between spontaneous herbivore species and cultivated plants under specific conditions of botanical garden greenhouses. In this study, the taxonomic and functional composition of sucking arthropod herbivore assemblages were correlated with leaf anatomical and surface features of 33 host species of Ficus L. (Moraceae) across four distantly located greenhouse complexes of botanical gardens. The analyses revealed that the species number and abundance of scale insects and their individual families, sessile phloem feeders, total phloem feeders and total herbivores were significantly positively correlated with the thickness of leaf lamina, epidermis and mesophyll, stomatal length and width, and the presence of abaxial multiple epidermis and weak ferruginous non-glandular trichomes. Significant negative correlations were revealed between the same herbivore parameters and the density of glandular trichomes. Heliomorphic leaves supported higher abundance and species richness of sessile phloem feeders compared to sciomorphic ones. The parameters of some phloem and mesophyll feeder taxa also correlated with non-glandular trichome length and density, type of trichomes and epicuticular wax layer, and the presence of calcium oxalate crystals in the epidermis. Results of the study suggest that the leaf architecture-related herbivory trends under greenhouse conditions are similar to those occurring in natural ecosystems when considering the functional significance of particular leaf traits, and remain relevant at the scale of particular plant taxa with disregard of spatial factor.