Site-directed mutagenesis of proline 94 to alanine in amicyanin converts a true electron transfer reaction into one that is kinetically coupled

Amicyanin is a type I copper protein that mediates electron transfer (ET) from methylamine dehydrogenase (MADH) to cytochrome c-551i. Pro(94) resides in the "ligand loop" of amicyanin, a sequence of amino acids that contains three of the four copper ligands. ET from the reduced O-quinol tryptophan tryptophylquinone of MADH to oxidized P94A amicyanin is a true ET reaction that exhibits values of electronic coupling (H(AB)) and reorganization energy (lambda) that are the same as for the reaction of native amicyanin. In contrast, the parameters for the ET reaction from reduced P94A amicyanin to oxidized cytochrome c-551i have been significantly altered as a consequence of the mutation. These values of H(AB) and lambda are 8.3 cm(-)(1) and 2.3 eV, respectively, compared to values of 0.3 cm(-)(1) and 1.2 eV for the reaction of native reduced amicyanin. The crystal structure of reduced P94A amicyanin exhibits two alternate conformations with the positions of the copper 1.4 A apart [Carrell, C. J., Sun, D., Jiang, S., Davidson, V. L., and Mathews, F. S. (2004) Biochemistry 43, 9372-9380]. In one of these, conformation B, a water molecule has replaced Met(98) as a copper ligand, and the ET distance to the heme of the cytochrome is increased by 1.4 A. Analysis of these structures suggests that the true k(ET) for ET from the copper in conformation B to heme would be much less than for ET from conformation A. A novel kinetic mechanism is proposed to explain these data in which the reduction of Cu(2+) by methylamine dehydrogenase is a true ET reaction while the oxidation of Cu(1+) by cytochrome c-551i is kinetically coupled ET. By comparison of the temperature dependence of the observed rate of the coupled ET reaction from reduced P94A amicyanin to cytochrome c-551i with the predicted rates and temperature dependence for the true ET reaction from conformation A, it was possible to determine the K(eq) and values of DeltaH degrees and DeltaS degrees that are associated with the non-ET reaction that modulates the observed ET rate.     

血管内皮细胞内皮素代谢的数值分析

将培养的人胚肾小球血管单层内皮细胞置于剪应力分别为6．5dyn/cm^2和13．0dyn/cm^2的定常层流中剪切10小时,样品中的内皮素（ET）分泌量用放射免疫法测定。结果表明,剪切应力和剪切作用时间对内皮细胞内皮素的代谢均有显著的影响,其影响不是简单的线性增减,而呈较复杂的非线性特征,内皮素累积含量和分泌速率,随剪切时间的变化的关系可用Logistic方程来描述,获得了反映内皮素（ET）分泌规律的大量特征方程和特征数,为了解体内发生于血管壁的病理生理过程中内皮素分泌规律提供了实验数据。     

Velocity profiles in stenosed tube models using magnetic resonance imaging

A time-of-flight MRI velocity measurement technique is evaluated against corresponding LDV measurements in a constriction tube model over a range of physiologic flow conditions. Results from this study show that MR displacement images can: 1) be obtained within both laminar and turbulent jets (maximum stenotic Re approximately equal to 4,200), 2) measure mean jet velocities up to 172 cm/s, and, 3) detect low forward and reverse stenosis (0 less than or equal to L/D less than or equal to 2). Regions between the jet termination point and re-establishment of laminar flow (Re greater than or equal to 1500, greater than or equal to 1000, and greater than or equal to 110 downstream of 40, 60, and 80 percent stenosis, respectively) cannot presently be detected by this technique.     

A numerical simulation of steady flow fields in a bypass tube.

Steady flow in a complete by-pass tube was simulated numerically. The study was to consider a complete flow field, which included both the by-pass and the host tubes. The changes of the hemodynamics were investigated with three parameters: the inlet flow Reynolds number (Re), anastomotic angle (alpha) and the position of the occlusion in the host tube. The baseline flow field was set up with Re=200, alpha=45 degrees and the centered position of occlusion. The parametric study was then conducted on combination of Re=100, 200, 400, alpha=35 degrees, 45 degrees, 60 degrees, 75 degrees, 90 degrees and three occlusion positions: left, center and right. It was found that in the baseline case, large slow/recirculation flows could be seen in the host tube both upstream and downstream of the occlusion. The separation points were on the opposite walls to the junctions. Recirculation zones were also found near the toe and in the proximal outer wall of the by-pass tube. Their sizes were about one diameter of the tube or smaller. In some cases, pairing vortices could be seen in the host tube upstream of the occlusion. The shear rate distribution associated with the flow fields was presented. The flow pattern obtained was agreeable to those observed experimentally by other investigators. The difference of the flow fields between a complete bypass and simple anastomosis was discussed. The present numerical code provides a preliminary simulation/design tool for bypass graft flows.     

Computational analysis of confined jet flow and mass transport in a blind tube.

A computational analysis of confined nonimpinging jet flow in a blind tube is performed as an initial investigation of the underlying fluid and mass transport mechanics of tracheal gas insufflation. A two-dimensional axisymmetric model of a laminar steady jet flow into a concentric blind-end tube is put forth and the governing continuity, momentum, and convection-diffusion equations are solved with a finite element code. The effects of the jet diameter based Reynolds number (Re(j)), the ratio of the jet-to-outer tube diameters (epsilon), and the Schmidt number (Sc) are evaluated with the determined velocity and contaminant concentration fields. The normalized penetration depth of the jet is found to increase linearly with increasing Re(j) for epsilon = O(0.1). For a given epsilon, a ring vortex that develops is observed to be displaced downstream and radially outward from the jet tip for increasing Re(j). The axial shear stress profile along the inside wall of the outer tube possesses regions of fixed shear stress in addition to a local minimum and maximum in the vicinity of the jet tip. Corresponding regions of axial shear stress gradients exist between the fixed shear stress regions and the local extrema. Contaminant concentration gradients develop across the ring vortex indicating the inward diffusion of contaminant into the jet flow. For fixed epsilon and Sc and Re(j) approximately 900, normalized contaminant flow rate is observed to be approximately twice that of simple diffusion. This model predicts modest net axial contaminant transport enhancement due to convection-diffusion interaction in the region of the ring vortex.     

Velocity field of pulsatile flow in a porous tube

This paper describes velocity fields for fully developed periodic laminar flow in a rigid tube with a porous wall. We obtained an analytical solution of the flow by the linear approximation of the Navier-Stokes equation. Unlike the previous works with a constant seepage rate along the axis, we used a wall velocity which contained hydraulic permeation constant Lp. The axial velocity profile shows a local maximum velocity near the wall at a large Womersley number alpha. This suggests that concentration polarization in porous tubular membrane may be reduced at high frequencies if a membrane device is operated under pulsatile flow conditions. The magnitude of wall permeation velocity decreases linearly along the tube axis because the damping of the pressure difference between the inside and the outside of the tube is very small.     

剪切流场中内皮细胞膜张应力累加效应的实验研究-离体血管段长度与其蛋白质代谢的相关性

用离体血管灌流试验验证冯元桢等关于血管内皮细胞的膜张应力逆血流方向轻加的理论分析。用０．１２Ｎ／ｍ＾２剪应力剪切人脐静脉段内皮细胞２０ｈ后,两种不同长度离体血管的内皮素平均分泌速率无显著差异,但在分泌速率的曲线形态,特征方程和最大分泌速率,最小分泌速率及分泌速率变异系数上差异较大;ＡＴⅡ的平均分泌速率差异彩显著,１１ｃｍ处理比２１ｃｍ处理高６８％。     

An apparatus to study the response of cultured endothelium to shear stress

An apparatus which has been developed to study the response of cultured endothelial cells to a wide range of shear stress levels is described. Controlled laminar flow through a rectangular tube was used to generate fluid shear stress over a cell-lined coverslip comprising part of one wall of the tube. A finite element method was used to calculate shear stresses corresponding to cell position on the coverslip. Validity of the finite element analysis was demonstrated first by its ability to generate correctly velocity profiles and wall shear stresses for laminar flow in the entrance region between infinitely wide parallel plates (two-dimensional flow). The computer analysis also correctly predicted values for pressure difference between two points in the test region of the apparatus for the range of flow rates used in these experiments. These predictions thus supported the use of such an analysis for three-dimensional flow. This apparatus has been used in a series of experiments to confirm its utility for testing applications. In these studies, endothelial cells were exposed to shear stresses of 60 and 128 dynes/cm2. After 12 hr at 60 dynes/cm2, cells became aligned with their longitudinal axes parallel to the direction of flow. In contrast, cells exposed to 128 dynes/cm2 required 36 hr to achieve a similar reorientation. Interestingly, after 6 hr at 128 dynes/cm2, specimens passed through an intermediate phase in which cells were aligned perpendicular to flow direction. Because of its ease and use and the provided documentation of wall shear stress, this flow chamber should prove to be a valuable tool in endothelial research related to atherosclerosis.     

AGS3, an alpha(1-3)glucan synthase gene family member of Aspergillus fumigatus, modulates mycelium growth in the lung of experimentally infected mice

The cell wall of human fungal pathogen Aspergillus fumigatus protects the fungus against threats from environment and interacts with the host immune system. Alpha(1-3)glucan is the major polysaccharide of Aspergillus fumigatus cell wall, and it has been shown to contribute to the virulence of diverse fungal pathogens. In A. fumigatus, three putative alpha(1-3)glucan synthase genes AGS1, AGS2 and AGS3 have been identified. AGS1 is responsible for cell wall alpha(1-3)glucan biosynthesis, but strains with deletions of either AGS1 or AGS2 are not defective in virulence [Beauvais, A., Maubon, D., Park, S., Morelle, W., Tanguy, M., Huerre, M., Perlin, D.S., Latgé, J. P., 2005. Two alpha(1-3) glucan synthases with different functions in Aspergillus fumigatus. Appl. Environ. Microbiol. 71, 1531-1538]. In contrast, we present evidence that AGS3 is also responsible for cell wall alpha(1-3)glucan biosynthesis and can modulate the virulence of A. fumigatus. An AGS3 deletion strain was found to produce faster and more robust disease than the parental strain in an experimental mouse model of aspergillosis. The apparent hyper-virulence in the AGS3-deleted mutant was correlated with an increased melanin content of the conidial cell wall, a better resistance to reactive oxygen species and a quicker germination rate. These results suggest an indirect role for AGS3 in virulence through an adaptive mechanism.     

Altered role of smooth muscle endothelin receptors in coronary endothelin-1 and alpha1-adrenoceptor-mediated vasoconstriction in Type 2 diabetes

Regulation of vascular tone and blood flow involves interactions between numerous local and systemic vascular control signals, many of which are altered by Type 2 diabetes (T2D). Vascular responses to endothelin-1 (ET-1) are mediated by endothelin type A (ET(A)) and type B (ET(B)) receptors that have been implicated in cross talk with alpha(1)-adrenoceptors (alpha(1)-AR). ET(A) and ET(B) receptor expression and plasma ET-1 levels are elevated in T2D; however, whether this influences coronary alpha(1)-AR function has not been examined. Therefore, we examined the effect of ET(A) and ET(B) receptor inhibition on coronary vasoconstriction to ET-1 and alpha(1)-AR activation in a mouse model of T2D. Coronary vascular responses were examined in isolated mouse hearts from control and diet-induced T2D C57BL/6J mice. Responses to ET-1 and the selective alpha(1)-AR agonist phenylephrine (PE) were examined alone and in the presence of the nitric oxide synthase inhibitor N(omega)-nitro-l-arginine methyl ester (l-NAME) alone or in combination with selective ET(A) or ET(B) receptor inhibitors BQ-123 and BQ-788, respectively. Vasoconstriction to ET-1 was enhanced, whereas ET(B), but not ET(A), receptor blockade reduced basal coronary tone in T2D hearts. In the presence of l-NAME, ET(A) receptor inhibition attenuated ET-1 vasoconstriction in both groups, whereas ET(B) inhibition abolished this response only in control hearts. In addition, ET(A) inhibition enhanced alpha(1)-AR-mediated vasoconstriction in T2D, but not control, hearts following l-NAME treatment. Therefore, in this model, enhanced coronary ET-1 responsiveness is mediated primarily through smooth muscle ET(B) receptors, whereas the interaction with alpha(1)-ARs is mediated solely through the ET(A) receptor subtype.     

Tube structures of probable microbial origin in the Neoarchean Carawine Dolomite, Hamersley Basin, Western Australia

The ~2.63 Ga Carawine Dolomite, Hamersley Basin, Western Australia, preserves tube structures of probable microbial origin that formed in a low energy environment. The tubes are 0.4–1.8 cm in diameter and at least 10–16 cm long in outcrop. The tubes are defined by dark, 45-µm-thick dolomicritic walls, whereas the tube fill and host rock are composed of 30 µm, cloudy dolomite crystals and rare 170- to 425-µm-wide, dark well-sorted clasts. Closely spaced, rarely discontinuous laminae coat the insides of tubes; less closely spaced, peaked, discontinuous laminae coat the outsides of tubes. The laminae on the outsides of tubes are often intercalated with mammilate structures. The presence of probable microbial coatings on both the insides and the outsides of the tube walls requires that the tubes formed above the sediment–water interface. These tube structures probably formed during gas-charged fluid escape, similar to tubes observed in ancient and modern hydrocarbon seeps and cylindrical water transfer structures in sandstones. The laminae that coat the tubes have very similar geometries to modern biofilms that form in both turbulent and laminar flow, and their geometries probably reflect flow conditions during the fluid escape. The identification of these structures suggests that the preserved interaction between fluid escape and microbial growth in carbonates may be more common than previously thought.     

A single methionine residue dictates the kinetic mechanism of interprotein electron transfer from methylamine dehydrogenase to amicyanin

Amicyanin is a type 1 copper protein that is the natural electron acceptor for the quinoprotein methylamine dehydrogenase (MADH). A P52G amicyanin mutation increased the Kd for complex formation and caused the normally true electron transfer (ET) reaction from O-quinol MADH to amicyanin to become a gated ET reaction (Ma, J. K., Carrell, C. J., Mathews, F. S., and Davidson, V. L. (2006) Biochemistry 45, 8284-8293). One consequence of the P52G mutation was to reposition the side chain of Met51, which is present at the MADH-amicyanin interface. To examine the precise role of Met51 in this interprotein ET reaction, Met51 was converted to Ala, Lys, and Leu. The Kd for complex formation of M51A amicyanin was unchanged but the experimentally determined electronic coupling increased from 12 cm-1 to 142 cm-1, and the reorganization energy increased from 2.3 to 3.1 eV. The rate and salt dependence of the proton transfer-gated ET reaction from N-quinol MADH to amicyanin is also changed by the M51A mutation. These changes in ET parameters and rates for the reactions with M51A amicyanin were similar to those caused by the P52G mutation and indicated that the ET reaction had become gated by a similar process, most likely a conformational rearrangement of the protein ET complex. The results of the M51K and M51L mutations also have consequences on the kinetic mechanism of regulation of the interprotein ET with effects that are intermediate between what is observed for the reaction of the native amicyanin and M51A amicyanin. These data indicate that the loss of the interactions involving Pro52 were primarily responsible for the change in Kd for P52G amicyanin, while the interactions involving the Met51 side chain are entirely responsible for the change in ET parameters and conversion of the true ET reaction of native amicyanin into a conformationally gated ET reaction.     

Characterization of the contractile and relaxant action of the endothelin-1 precursor, big endothelin-1, in the isolated rat basilar artery

The presence of functional endothelin converting enzyme (ECE) activity in basilar artery ring segments was investigated by measuring the contractile and relaxant effects of big endothelin (ET)-1. Under resting tension conditions cumulative application of big ET1-1 elicited a concentration-related contraction with the concentration-effect curve (CEC) shifted to the right against ET-1 by a factor of 31 and 29 in segments with the endothelium intact or mechanically removed, respectively. Preincubation with the ET(A) receptor antagonist, BQ123, induced an apparently parallel rightwards shift without affecting the maximum contraction. This shift was more pronounced for ET-1 than for big ET-1. With the putative ECE inhibitor phosphoramidon (10(-3) M) in the bath a small rightwards shift of the CEC for big ET-1 was observed in control segments and a more marked one in de-endothelialized segments. In segments precontracted with prostaglandin (PG) F(2alpha) big ET-1 induced a significant although transient relaxation whereas ET-1 did not. However, in the presence of BQ123 both ET-1 and big ET-1 elicited concentration-related relaxation with a significantly higher maximum effect obtained with big ET-1. The potency was 13 fold higher for ET-1, which is markedly less than that found for contraction. The results, therefore, suggest 1) the presence of functional ECE-activity in the rat basilar artery wall, and 2) differences in the functional ECE activity located in the endothelium and media.     

Re-engineering monovalent cation binding sites of methylamine dehydrogenase: effects on spectral properties and gated electron transfer

Methylamine dehydrogenase (MADH) is a tryptophan tryptophylquinone (TTQ)-dependent enzyme that catalyzes the oxidative deamination of primary amines. Monovalent cations are known to affect the spectral properties of MADH and to influence the rate of the gated electron transfer (ET) reaction from substrate-reduced MADH to amicyanin. Two putative monovalent cation binding sites in MADH have been identified by X-ray crystallography [Labesse, G., Ferrari, D., Chen, Z.-W., Rossi, G.-L., Kuusk, V., McIntire, W. S., and Mathews, F. S. (1998) J. Biol. Chem. 273, 25703-25712]. One requires cation-pi interactions involving residue alpha Phe55. An alpha F55A mutation differentially affects these two monovalent cation-dependent phenomena. The apparent K(d) associated with spectral perturbations increases 10-fold. The apparent K(d) associated with enhancement of the gated ET reaction becomes too small to measure, indicating that either it has decreased more than 1000-fold or the mutation has caused a conformational change that eliminates the requirement for the cation for the gated ET. These results show that of the two binding sites revealed in the structure, cation binding to the distal site, which is stabilized by the cation-pi interactions, is responsible for the spectral perturbations. Cation binding to the proximal site, which is stabilized by several oxygen ligands, is responsible for the enhancement of the rate of gated ET. Another site-directed mutant, alpha F55E MADH, exhibited cation binding properties that were the same as those of the native enzyme, indicating that interactions with the carboxylate of Glu can effectively replace the cation-pi interactions with Phe in stabilizing monovalent cation binding to the distal site.     

J1 acylase, a glutaryl-7-aminocephalosporanic acid acylase from Bacillus laterosporus J1, is a member of the alpha/beta-hydrolase fold superfamily

J1 acylase, a glutaryl-7-aminocephalosporanic acid acylase (GCA) isolated from Bacillus laterosporus J1, has been conventionally grouped as the only member of class V GCA, although its amino acid sequence shares less than 10% identity with members of other classes of GCA. Instead, it shows higher sequence similarities with Rhodococcus sp. strain MB1 cocaine esterase (RhCocE) and Acetobacter turbidans alpha-amino acid ester hydrolase (AtAEH), members of the alpha/beta-hydrolase fold superfamily. Homology modeling and secondary structure prediction indicate that the N-terminal region of J1 acylase has an alpha/beta-hydrolase folding pattern. The catalytic triads in RhCocE and AtAEH were identified in J1 acylase as S125, D264 and H309. Mutations to alanine at these positions were found to completely inactivate the enzyme. These results suggest that J1 acylase is a member of the alpha/beta-hydrolase fold superfamily with a serine-histidine-aspartate catalytic triad.     

A1 adenosine receptors of bovine brain couple to guanine nucleotide-binding proteins Gi1, Gi2, and Go

A1 adenosine receptors and associated guanine nucleotide-binding proteins (G proteins) were purified from bovine cerebral cortex by affinity chromatography (Munshi, R., and Linden, J. (1989) J. Biol. Chem. 264, 14853-14859). In this study we have identified the pertussis toxin-sensitive G protein subunits that co-purify with A1 adenosine receptors by immunoblotting with specific antipeptide antisera. Gi alpha 1, Gi alpha 2, Go alpha, G beta 35, and G beta 36 were detected. Of the total [35S]guanosine 5'-O-(3-thio)triphosphate [( 35S]GTP gamma S) binding sites, Gi alpha 1 and Go alpha each accounted for greater than 37% whereas Gi alpha 2 comprised less than 13%. G beta 35 was found in excess over G beta 36. Low molecular mass (21-25 kDa) GTP-binding proteins were not detected. We also examined the characteristics of purified receptors and various purified bovine brain G proteins reconstituted into phospholipid vesicles. All three alpha-subunits restored GTP gamma S-sensitive high affinity binding of the agonist 125I-aminobenzyladenosine to a fraction (25%) of reconstituted receptors with a selectivity order of Gi2 greater than Go greater than or equal to Gi1 (ED50 values of G proteins measured as fold excess over the receptor concentration were 4.7 +/- 1.2, 24 +/- 5, and 34 +/- 7, respectively). Furthermore, receptors occupied with the agonist R-phenylisopropyladenosine catalytically increased the rate of binding of [35S]GTP gamma S to reconstituted G proteins by 6.5-8.5-fold. These results suggest that A1 adenosine receptors couple indiscriminately to pertussis toxin-sensitive G proteins.     

DREB1A promotes root development in deep soil layers and increases water extraction under water stress in groundnut

Water deficit is a major yield‐limiting factor for many crops, and improving the root system has been proposed as a promising breeding strategy, although not in groundnut (Arachis hypogaea L.). The present work was carried out mainly to assess how root traits are influenced under water stress in groundnut, whether transgenics can alter root traits, and whether putative changes lead to water extraction differences. Several transgenic events, transformed with DREB1A driven by the rd29 promoter, along with wild‐type JL24, were tested in a lysimeter system that mimics field conditions under both water stress (WS) and well‐watered (WW) conditions. The WS treatment increased the maximum rooting depth, although the increase was limited to about 20% in JL24, compared to 50% in RD11. The root dry weight followed a similar trend. Consequently, the root dry weight and length density of transgenics was higher in layers below 100‐cm depth (Exp. 1) and below 30 cm (Exp. 2). The root diameter was unchanged under WS treatment, except a slight increase in the 60–90‐cm layer. The root diameter increased below 60 cm in both treatments. In the WW treatment, total water extraction of RD33 was higher than in JL24 and other transgenic events, and somewhat lower in RD11 than in JL24. In the WS treatment, water extraction of RD2, RD11 and RD33 was higher than in JL24. These water extraction differences were mostly apparent in the initial 21 days after treatment imposition and were well related to root length density in the 30–60‐cm layer (R² = 0.68), but not to average root length density. In conclusion, water stress promotes rooting growth more strongly in transgenic events than in the wild type, especially in deep soil layers, and this leads to increased water extraction. This opens an avenue for tapping these characteristics toward the improvement of drought adaptation in deep soil conditions, and toward a better understanding of genes involved in rooting in groundnut.     

A pyrrolidine-based specific inhibitor of cytosolic phospholipase A(2)alpha blocks arachidonic acid release in a variety of mammalian cells

We analyzed a recently reported (K. Seno, T. Okuno, K. Nishi, Y. Murakami, F. Watanabe, T. Matsuur, M. Wada, Y. Fujii, M. Yamada, T. Ogawa, T. Okada, H. Hashizume, M. Kii, S.-H. Hara, S. Hagishita, S. Nakamoto, J. Med. Chem. 43 (2000)) pyrrolidine-based inhibitor, pyrrolidine-1, against the human group IV cytosolic phospholipase A(2) alpha-isoform (cPLA(2)alpha). Pyrrolidine-1 inhibits cPLA(2)alpha by 50% when present at approx. 0.002 mole fraction in the interface in a number of in vitro assays. It is much less potent on the cPLA(2)gamma isoform, calcium-independent group VI PLA(2) and groups IIA, X, and V secreted PLA(2)s. Pyrrolidine-1 blocked all of the arachidonic acid released in Ca(2+) ionophore-stimulated CHO cells stably transfected with cPLA(2)alpha, in zymosan- and okadaic acid-stimulated mouse peritoneal macrophages, and in ATP- and Ca(2+) ionophore-stimulated MDCK cells.     

Evapotranspiration at the land/water interface in a semi-arid drainage basin

1. Evapotranspiration (ET) is a major source of water depletion from riverine systems in arid and semiarid climates. Water budgets have produced estimates of total depletions from riparian vegetation ET for a 320‐km reach of the Middle Rio Grande, New Mexico, U.S.A., that have ranged from 20 to 50% of total depletions from the river. 2. Tower‐based micrometeorological measurements of riparian zone ET throughout the growing season using three‐dimensional eddy covariance provided high quality estimates of ET at the stand scale. 3. A dense stand of salt cedar (111–122 cm year^–1) and a mature cottonwood (Populus deltoides ssp. wislizenia Eckenwelder) stand with an extensive understory of salt cedar (Tamaria ramosissima Ledeb) and Russian olive (Eleagnus angustifolia L.) (123 cm year^–1) had the highest rates of annual ET. A mature cottonwood stand with a closed canopy had intermediate rates of ET (98 cm year^–1). A less dense salt cedar stand had the lowest rates of ET (74–76 cm year^–1). 4. Summer leaf area index (LAI) measurements within the four stands were positively correlated with daily ET rates. LAI measurements throughout the growing season coupled to riparian vegetation classification is a promising method for improving riverine corridor estimates of total annual riparian zone ET along a reach of river. 5. Combining recent estimates of the extent of riparian vegetation along the 320 km length of the Middle Rio Grande, from Landsat 7 imagery with annual growing season measurements of ET at the four riparian stands yields a first‐order riverine corridor estimate of total riparian zone ET of 150–250 × 10⁶ m³ year^–1. This is approximately 20–33% of total estimated depletions along this reach of river.     

Exaggerated hypoxic pulmonary hypertension in endothelin B receptor-deficient rats

Mechanisms by which endothelin (ET)-1 mediates chronic pulmonary hypertension remain incompletely understood. Although activation of the ET type A (ET(A)) receptor causes vasoconstriction, stimulation of ET type B (ET(B)) receptors can elicit vasodilation or vasoconstriction. We hypothesized that the ET(B) receptor attenuates the development of hypoxic pulmonary hypertension and studied a genetic rat model of ET(B) receptor deficiency (transgenic sl/sl). After 3 wk of severe hypoxia, the transgenic sl/sl pulmonary vasculature lacked expression of mRNA for the ET(B) receptor and developed exaggerated pulmonary hypertension that was characterized by elevated pulmonary arterial pressure, diminished cardiac output, and increased total pulmonary resistance. Plasma ET-1 was fivefold higher in transgenic sl/sl rats than in transgenic controls. Although mRNA for prepro-ET-1 was not different, mRNA for ET-converting enzyme-1 was higher in transgenic sl/sl than in transgenic control lungs. Hypertensive lungs of sl/sl rats also produced less nitric oxide metabolites and 6-ketoprostaglandin F(1alpha), a metabolite of prostacyclin, than transgenic controls. These findings suggest that the ET(B) receptor plays a protective role in the pulmonary hypertensive response to chronic hypoxia.