ATP synthesis without R210 of subunit a in the Escherichia coli ATP synthase

Interactions between subunit a and oligomeric subunit c are essential for the coupling of proton translocation to rotary motion in the ATP synthase. A pair of previously described mutants, R210Q/Q252R and P204T/R210Q/Q252R [L.P. Hatch, G.B. Cox and S.M. Howitt, The essential arginine residue at position 210 in the a subunit of the Escherichia coli ATP synthase can be transferred to position 252 with partial retention of activity, J. Biol. Chem. 270 (1995) 29407-29412] has been constructed and further analyzed. These mutants, in which the essential arginine of subunit a, R210, was switched with a conserved glutamine residue, Q252, are shown here to be capable of both ATP synthesis by oxidative phosphorylation, and ATP-driven proton translocation. In addition, lysine can replace the arginine at position 252 with partial retention of both activities. The pH dependence of ATP-driven proton translocation was determined after purification of mutant enzymes, and reconstitution into liposomes. Proton translocation by the lysine mutant, and to a lesser extent the arginine mutant, dropped off sharply above pH 7.5, consistent with the requirement for a positive charge during function. Finally, the rates of ATP synthesis and of ATP-driven proton translocation were completely inhibited by treatment with DCCD (N,N'-dicyclohexylcarbodiimide), while rates of ATP hydrolysis by the mutants were not significantly affected, indicating that DCCD modification disrupts the F(1)-F(o) interface. The results suggest that minimal requirements for proton translocation by the ATP synthase include a positive charge in subunit a and a weak interface between subunit a and oligomeric subunit c.     

Latitudinal patterns of magnitude and interannual variability in net ecosystem exchange regulated by biological and environmental variables

Over the last two and half decades, strong evidence showed that the terrestrial ecosystems are acting as a net sink for atmospheric carbon. However the spatial and temporal patterns of variation in the sink are not well known. In this study, we examined latitudinal patterns of interannual variability (IAV) in net ecosystem exchange (NEE) of CO₂ based on 163 site-years of eddy covariance data, from 39 northern-hemisphere research sites located at latitudes ranging from ∼29°N to ∼64°N. We computed the standard deviation of annual NEE integrals at individual sites to represent absolute interannual variability (AIAV), and the corresponding coefficient of variation as a measure of relative interannual variability (RIAV). Our results showed decreased trends of annual NEE with increasing latitude for both deciduous broadleaf forests and evergreen needleleaf forests. Gross primary production (GPP) explained a significant proportion of the spatial variation of NEE across evergreen needleleaf forests, whereas, across deciduous broadleaf forests, it is ecosystem respiration (Re). In addition, AIAV in GPP and Re increased significantly with latitude in deciduous broadleaf forests, but AIAV in GPP decreased significantly with latitude in evergreen needleleaf forests. Furthermore, RIAV in NEE, GPP, and Re appeared to increase significantly with latitude in deciduous broadleaf forests, but not in evergreen needleleaf forests. Correlation analyses showed air temperature was the primary environmental factor that determined RIAV of NEE in deciduous broadleaf forest across the North American sites, and none of the chosen climatic factors could explain RIAV of NEE in evergreen needleleaf forests. Mean annual NEE significantly increased with latitude in grasslands. Precipitation was dominant environmental factor for the spatial variation of magnitude and IAV in GPP and Re in grasslands.     

The gross anatomy,cellular structure,and fatty acid composition of adipose tissue in captive polar bears (Ursus maritimus)

The gross anatomy, microscopic structure, and fatty acid composition of adipose tissue in four elderly polar bears (Ursus maritimus), that had each been in captivity for more than 24 years, were compared with that of young wild specimens. Total dissections were performed on two specimens. In both, the proportion of adipose tissue in the body was similar to that of younger wild bears, but the total adipocyte complement was smaller than that of the wild bears in one specimen, and within the normal range in the other. There was proportionately more intra-abdominal adipose tissue, particularly in the inner ventral wall of abdomen depot, in which the number of adipocytes was substantially greater than expected. The fatty acid composition of the triacylglycerols differed substantially from that of the wild bears, reflecting their diet of fish, domesticated herbivores, bread, and fruit, but there were also significant differences between cage-mates eating similar diets. © 1993 Wiley-Liss, Inc.     

Water relations parameters and tissue development in somatic and zygotic embryos of three pinaceous conifers

There is increasing interest in using somatic embryogenesis to meet the demand for high quality seedlings. However, in vitro production of propagules on a large scale depends on the optimization of the maturation and germination steps promoted by desiccation and subsequent imbibition of the embryo, respectively. It is therefore important to characterize zygotic and somatic embryos in terms of their water relations. Bound water, elastic modulus, osmotic potential at full turgor, and relative water content at turgor loss point were determined for somatic and zygotic embryos of western larch and loblolly pine and somatic embryos of white spruce at two developmental stages. These water relations parameters were derived from water-release curves obtained by suspending tissue samples in sealed vials over unsaturated NaCl solutions of known water potential. There was little difference in water relations parameters among species but marked stage dependency for bound water and elastic modulus. The amount of bound water was lowest in western larch somatic embryos (0.02-0.07) and highest in zygotic loblolly pine embryos (0.10-0.18). Elastic modulus ranged from 2.5 to 6.2 MPa in somatic embryos but varied between 1.4 and 1.8 MPa in zygotic embryos. The osmotic potential was lower in somatic embryos than in their zygotic counterparts. Our results show that water relations parameters are remarkably conservative across species but that, within a given species, these parameters are stage specific. It would seem, therefore, that desiccation protocols might be developed for a given developmental stage and applied across a range of species without the confounding effects of differences in water relations parameters.     

Stable-Carbon-Isotope Composition of Fatty Acids in Hydrothermal Vent Mussels Containing Methanotrophic and Thiotrophic Bacterial Endosymbionts

Fatty acid biomarker analysis coupled with gas chromatography-isotope ratio mass spectrometry was used to confirm the presence of methanotrophic and thiotrophic bacterial endosymbionts in the tissues of a hydrothermal vent mussel (Bathymodiolus sp.), collected from the Menez Gwen vent field on the mid-Atlantic ridge. Monounsaturated (n-8) fatty acids, which are diagnostic of methanotrophic bacteria, were detected in all three types of tissues examined (gill, posterior adductor, and mantle), although levels were highest in gill tissues where the bacteria were found. Stable-carbon-isotope compositions (δ-¹³C per mille relative to that of Peedee belemnite) of fatty acids for all three tissues ranged from −24.9 to −34.9‰, which encompasses the range predicted for both thiotroph- and methanotroph-based nutrition. The data suggest that these thio- and methanotrophic bacterial endosymbionts are equally important in the nutrition of the vent mussel at this particular vent site.     

Effect of dietary Ca and Cd level of pregnant rats on reproduction and on dam and progeny tissue mineral concentrations.

Pregnant Sprague-Dawley rats were used to determine the effects of the addition of 200 ppm of Cd (as CdCl2) to the diet factorially with two levels of dietary Ca (0.07% and 0.96%) on reproductive performance, concentrations of Cd, Cu, Fe, Zn, Ca and Mg in dam liver and kidney and in newborn progeny. High Cd significantly increased liver and kidney Cd, Zn and Ca and decreased liver Fe. High dietary Ca partially protected against accumulation of Cd in liver and kidney but had no effect on concentration of other elements. Number of live or stillborn pups per litter was not significantly affected by diet but high Cd significantly reduced pup birth weight. No grossly abnormal pups were noted. Concentration of Cd in bodies of newborn pups was increased approximately 8.6-fold by high Cd in the diet of dams fed the 0.07% Ca-diet and 3.8-fold by high-Cd in the diet of dams fed the 0.96% Ca diet. Pup, Zn, Cu and Fe contents were significantly decreased and Ca was significantly increased by high-Cd in the maternal diet whereas pup Mg content was unchanged. Maternal Ca intake had no effect on concentration of Zn, Cu, Fe or Ca in newborn pups. The biological importance of the alteration in maternal and fetal tissue concentration of Zn, Cu and Fe by high-Cd maternal diets is unknown.     

Sarcocystis in free-ranging herbivores on the National Bison Range.

Heart, esophagus, diaphragm and skeletal muscle obtained from various herbivores on the National Bison Range were examined grossly for Sarcocystis. Sarcocystis was found in 81, 50, 50, and 13% of the mule deer, (Odocoileus hemionus), white-tailed deer (O. virginianus), elk (Cervus elaphus), and bison (Bison bison), respectively.