Responses in stomatal conductance to elevated CO2 in 12 grassland species that differ in growth form

Responses in stomatal conductance (g _st ) and leaf xylem pressure potential ( _leaf ) to elevated CO₂ (2x ambient) were compared among 12 tallgrass prairie species that differed in growth form and growth rate. Open-top chambers (OTCs, 4.5 m diameter, 4.0 m in height) were used to expose plants to ambient and elevated CO₂ concentrations from April through November in undisturbed tallgrass prairie in NE Kansas (USA). In June and August, _leaf was usually higher in all species at elevated CO₂ and was lowest in adjacent field plots (without OTCs). During June, when water availability was high, elevated CO₂ resulted in decreased g _st in 10 of the 12 species measured. Greatest decreases in g _st (ca. 50%) occurred in growth forms with the highest potential growth rates (C₃ and C₄ grasses, and C₃ ruderals). In contrast, no significant decrease in g _st was measured in the two C₃ shrubs. During a dry period in September, reductions in g _st at elevated CO₂ were measured in only two species (a C₃ ruderal and a C₄ grass) whereas increased g _st at elevated CO₂ was measured in the shrubs and a C₃ forb. These increases in g _st were attributed to enhanced _leaf in the elevated CO₂ plants resulting from increased soil water availability and/or greater root biomass. During a wet period in September, only reductions in g _st were measured in response to elevated CO₂. Thus, there was significant interspecific variability in stomatal responses to CO₂ that may be related to growth form or growth rate and plant water relations. The effect of growth in the OTCs, relative to field plants, was usually positive for g _st and was greatest (>30%) when water availability was low, but only 6–12% when _leaf was high.The results of this study confirm the importance of considering interactions between indirect effects of high CO₂ of plant water relations and direct effects of elevated CO₂ on g _st , particularly in ecosystems such as grasslands where water availability often limits productivity. A product of this interaction is that the potential exists for either positive or negative responses in g _st to be measured at elevated levels of CO₂.     

Atomic force microscope measurements and manipulation of Langmuir-Blodgett films with modified tips.

A simple method for rendering atomic force microscope tips and cantilevers hydrophilic or hydrophobic through glow discharge in an appropriate gas atmosphere is introduced. Force curves at different humidities of these modified cantilevers were taken on freshly cleaved mica (hydrophilic surface) and on a monolayer of dipalmitoylphosphatidylethanolamine transferred onto mica (hydrophobic surface) to characterize the behavior of the cantilevers on hydrophilic and hydrophobic surfaces. Furthermore, Langmuir-Blodgett bilayers, with a dipalmitoylphosphatidylethanolamine bottom layer and a dipalmitoylphosphatidylcholine top layer, were imaged in the constant force mode in a multimode atomic force microscope in air under controlled humidity conditions. The friction and elasticity signal were recorded parallel to the topography. By varying the force exerted by the tip on the sample, different layers of the Langmuir-Blodgett system could be removed or flattened. Removal exposed underlying layers that exhibited a different friction and elasticity behavior. Furthermore, force scans with tips rendered hydrophobic were taken on the different layers of the sample to characterize the hydrophilic/hydrophobic nature of the layers. Only by combining the results obtained by the different methods can the structure of the lipid layer systems be identified.     

Solid-state fermentation of carob pods byAspergillus niger for protein production: effect of particle size

Two strains ofAspergillus niger were cultured in solid-state fermentation system on carob pods ground from 1.25 to 8 mm diam. A particle size of 2.5 mm gave the highest protein content of the final product (20%, w/w) and 52% of the total soluble carbohydrates were utilized. The total tannin concentration of the carob pods decreased by 83% in 4 days of fermentation.T. Smail and O. Salhi are with the Laboratory of Microbiology, U.R.B.A.F., Institute of Biology, Tizi-Ouzou University, Algeria. J.S. Knapp is with the Department of Microbiology, The University of Leeds, Leeds LS2 9JT, UK;     

Characterization of the spontaneous murine B cell leukemia (BCL1). III. Evidence for monoclonality by using an anti-idiotype antibody.

We have raised an anti-idiotypic antibody against the cell surface IgM of the murine BCL1 tumor cells. This antiserum reacts exclusively with the IgM expressed on the tumor cells and detects a unique population of cells in the spleen and blood of the tumor-bearing mice. When these cells are stimulated in vitro with LPS, they secrete an IgM bearing the same idiotype as the cell surface Ig. These results are discussed in terms of a model for the immunotherapy of a chronic lymphocytic leukemia-like syndrome in mice.     

Catalytic detection principle for high-performance liquid chromatography: Determination of enantiomeric iodinated thyronines in blood serum

A method for the trace determination of iodinated thyronines with differentiation of the optical isomers by high performance liquid chromatography (HPLC) is described. The detection is effected by means of a catalytic principle based on the iodide-catalysed reaction of chloramine-T and N,N′-tetramethyldiaminodiphenylmethane, producing a coloured complex that can be measured spectrophotometrically at 600 nm. Owing to the selectivity of the catalytic reaction, iodine-containing compounds can be easily determined in a complex matrix such as blood plasma. The sensitivity is sufficient for the detection of plasma levels of iodinated thyronines. The limit of detection for thyroxine is in the sub-nanogram range. The enantiomers of thyronines can be separated on commercial reversed phases after pre-column synthesis of diastereomers. For this derivatization the reagent tert.-butyloxy-carbonyl-l-leucine-N-hydroxysuccinimide ester is used. The coupling of the stereospecific HPLC separation with the catalytic detector offers the possibility of determining both d- and l-thyroxine in human plasma.     

Matrix-assisted laser desorption mass spectrometry of rhodopsin and bacteriorhodopsin.

Matrix-assisted laser desorption ionization (MALDI) mass spectrometry has been used to obtain accurate molecular weight information for the integral membrane proteins bacteriorhodopsin and bovine rhodopsin desorbed from solubilized membrane preparations. Mass differences in the molecular weights measured for bleached and unbleached bacteriorhodopsin and rhodopsin indicate the removal of the retinal chromophores upon bleaching. The MALDI technique was also successful for determination of the major cleavage products obtained upon treatment of membrane bound rhodopsin with endoproteinase Asp-N and thermolysin. Our results indicate that the MALDI method is a useful means of obtaining accurate molecular weight information on hydrophobic proteins isolated in their native membranes.     

Hirudisins. Hirudin-derived thrombin inhibitors with disintegrin activity.

Recombinant hirudin variants have been designed which inhibit alpha-thrombin by the hirudin mechanism and which in addition exhibit disintegrin activity. These proteins, called "hirudisins," have been engineered by replacing the Ser-Asp-Gly-Glu sequence at the tip of hirudin's finger-like structure (residues 32-35) by Arg-Gly-Asp-Ser (RGDS) to yield hirudisin and Lys-Gly-Asp-Ser (KGDS) to obtain hirudisin-1. Comparison of thrombin inhibition activities showed that hirudisin is 2-fold more potent (K(i) = 160 +/- 70 fM) than hirudisin-1 (K(i) = 370 +/- 44 fM) and recombinant (r)-hirudin (K(i) = 270 +/- 50 fM). alpha-Thrombin-stimulated platelet aggregation was effectively inhibited by r-hirudin, hirudisin, and hirudisin-1 with IC50 of 5.7 to 6.8 nM. Unlike r-hirudin, hirudisin inhibits ADP-induced platelet aggregation (IC50 = 65 microM) 3- to 5-fold stronger than the linear GRGDS- and RGDS-peptide. Direct interaction of hirudisin with purified glycoprotein IIb-IIIa demonstrated that antiplatelet aggregation activity is due to the integrin-directed RGD motif. Disintegrin activity of hirudisin relative to that of reduced and carboxymethylated hirudisin suggests that the conformational strain favors binding to integrins. On the basis of these results, hirudisins appear to be interesting molecules for the design of potential antithrombotic agents with antithrombin as well as antiplatelet aggregation activities.     

IgM and IgG but not cytokine secretion is restricted to the CD27+ B lymphocyte subset.

In a recent study we reported that CD27 is expressed on a subpopulation of human B lymphocytes and presented circumstantial phenotypic evidence that CD27 expression may be acquired late during B cell differentiation. Here we present functional data showing that, after in vitro stimulation, CD27+ but not CD27- B cells secrete large amounts of both IgM and IgG. Using double immunofluorescence staining of CD27 and IgD, three functionally different B cell subsets representing distinct stages of B cell differentiation can be isolated: 1) the CD27- IgD+ B cells, which do not secrete appreciable Ig; 2) the CD27+IgD+ B cells, which exclusively secrete IgM; and 3) the CD27+IgD- B cells, which comprise the IgG-producing cells. Furthermore, costimulation of CD27- B cells with low m.w. B cell growth factor, in the presence or in the absence of a CD40 mAb, does not induce these cells to become Ig-secreting cells. Although CD27- B cells hardly secrete Ig of any isotype in response to Staphylococcus aureus+IL-2, these cells proliferate vigorously and express the IL-2R alpha chain (CD25) under these stimulatory conditions. Furthermore, both CD27- and CD27+ B cells are capable of producing similar amounts of IL-6 and TNF-alpha. Taken together, these findings indicate that CD27 is a unique non-Ig surface marker discriminating naive from primed B lymphocytes. Furthermore, the capacity to proliferate and to secrete the B cell differentiation factors IL-6 and TNF-alpha already exists at an early B cell differentiation stage at which the cells lack CD27 expression and are not induced to produce Ig.     

Synthesis of [D-Ala2,4'-125I-Phe3,Glu4]deltorphin and characterization of its delta opioid receptor binding properties.

Both [D-Ala2,Glu4]Deltorphin and [D-Ala2,4'-I-Phe3,Glu4]Deltorphin are highly selective ligands for delta, relative to mu, opioid receptors. Radiolabeled [D-Ala2, 4'-125I-Phe3,Glu4]Deltorphin ([125I]Deltorphin) was prepared with a specific activity of 2200 Ci/mmol from [D-Ala2, 4'-NH2-Phe3, Glu4]Deltorphin through a diazonium salt intermediate. The inhibition of [125I]Deltorphin binding to rat brain membranes by ligands selective for mu, delta, and kappa opioid receptors is consistent with binding by the radioligand to a single site having the properties of a delta opioid receptor. The results of these studies are in good agreement with those obtained by structurally different delta opioid receptor ligands. The similarity between the delta receptor site labeled by [125I]Deltorphin and those labeled by other delta receptor agonists, in contrast to differences seen by in vivo studies of their analgesic effects, is discussed.