Microbial dynamics during the decline of a spring diatom bloom in the Northeast Atlantic

The microbial dynamics during a spring diatom bloom declinewas monitored in the Northeast Atlantic during a 5-day Lagrangianstudy (8–12 April 2002). Phytoplankton abundance, compositionand health status were related to viral and bacterial abundance,zooplankton abundance and grazing rates, as well as bacterialproduction. Phytoplankton reached maximum concentration on Day3 (Chl a >5 µg L^–1) and declined on Day 5 (Chla 2 µg L^–1) and was dominated (70% of Chl a) bydiatoms. Bacterial production increased substantially to >20µg C L^–1 day^–1 on Day 3 and concomitantlylarge viruses decreased in number by half to <10 x 10³ mL^–1.This was followed by a 5-fold increase in large viruses on Day5, indicating infection and subsequent lysis on Days 3 and 5,respectively. Micro- and mesozooplankton grazing were not theprincipal cause for the decline of the bloom and pheophorbide-ashowing little variation in concentration from Days 1–4(100 ng L^–1) although doubled on Day 5. The poor physiologicalstatus of the diatoms, indicated by the high chlorophyllide-aconcentrations (50–480 ng L^–1), likely promoteda series of closely interrelated events involving bacteria andviruses leading to the demise of the diatom bloom.     

Functional characterization of voltage-gated K+ channels in mouse pulmonary artery smooth muscle cells

Mice are useful animal models to study pathogenic mechanisms involved in pulmonary vascular disease. Altered expression and function of voltage-gated K⁺ (K_V) channels in pulmonary artery smooth muscle cells (PASMCs) have been implicated in the development of pulmonary arterial hypertension. K_V currents (I_K(V)) in mouse PASMCs have not been comprehensively characterized. The main focus of this study was to determine the biophysical and pharmacological properties of I_K(V) in freshly dissociated mouse PASMCs with the patch-clamp technique. Three distinct whole cell I_K(V) were identified based on the kinetics of activation and inactivation: rapidly activating and noninactivating currents (in 58% of the cells tested), rapidly activating and slowly inactivating currents (23%), and slowly activating and noninactivating currents (17%). Of the cells that demonstrated the rapidly activating noninactivating current, 69% showed I_K(V) inhibition with 4-aminopyridine (4-AP), while 31% were unaffected. Whole cell I_K(V) were very sensitive to tetraethylammonium (TEA), as 1 mM TEA decreased the current amplitude by 32% while it took 10 mM 4-AP to decrease I_K(V) by a similar amount (37%). Contribution of Ca²⁺-activated K⁺ (K_Ca) channels to whole cell I_K(V) was minimal, as neither pharmacological inhibition with charybdotoxin or iberiotoxin nor perfusion with Ca²⁺-free solution had an effect on the whole cell I_K(V). Steady-state activation and inactivation curves revealed a window K⁺ current between –40 and –10 mV with a peak at –31.5 mV. Single-channel recordings revealed large-, intermediate-, and small-amplitude currents, with an averaged slope conductance of 119.4 ± 2.7, 79.8 ± 2.8, 46.0 ± 2.2, and 23.6 ± 0.6 pS, respectively. These studies provide detailed electrophysiological and pharmacological profiles of the native K_V currents in mouse PASMCs. K_V channels     

Reduced sickle erythrocyte dehydration in vivo by endothelin-1 receptor antagonists

Elevated plasma levels of cytokines such as endothelin-1 (ET-1) have been shown to be associated with sickle cell disease (SCD). However, the role of ET-1 in the pathophysiology of SCD is not entirely clear. I now show that treatment of SAD mice, a transgenic mouse model of SCD, with BQ-788 (0.33 mg·kg^–1·day^–1 intraperitoneally for 14 days), an ET-1 receptor B (ET_B) antagonist, induced a significant decrease in Gardos channel activity (1.7 ± 0.1 to 1.0 ± 0.4 mmol·10¹³ cell^–1·h^–1, n = 3, P = 0.019) and reduced the erythrocyte density profile by decreasing the mean density (D₅₀; n = 4, P = 0.012). These effects were not observed in mice treated with BQ-123, an ET-1 receptor A (ET_A) antagonist. A mixture of both antagonists induced a similar change in density profile as with BQ-788 alone that was associated with an increase in mean cellular volume and a decrease in corpuscular hemoglobin concentration mean. I also observed in vitro effects of ET-1 on human sickle erythrocyte dehydration that was blocked by BQ-788 and a mixture of ET_B/ET_A antagonists but not by ET_A antagonist alone. These results show that erythrocyte hydration status in vivo is mediated via activation of the ET_B receptor, leading to Gardos channel modulation in SCD. cellular dehydration; Gardos channel; transgenic sickle mice     

The Effect of Mutations in the T-DNA Encoded Auxin Pathway on the Endogenous Phytohormone Content in Cloned Nicotiana tabacum Crown Gall Tissues

We analyzed the endogenous auxin and cytokinin levels of clonedNicotiana tabacum SR 1-lines induced either by the wild-typeAgrobacterium tumefaciens C58 strain or by mutants affectedin the T-DNA-encoded IAA biosynthesis pathway. The wild-typeSR1-C58 line contained up to 20 times more IAA than a nontransformedSRI-callus line. The mutant lines affected in gene 1 (iaaM^–)or gene 2 (iaaH^–) contained intermediate levels of IAA. Analysis of the endogenous levels of indole-3-acetamide (IAM)in the nontransformed SR 1 callus line, the wild-type SR1-C58and the two mutant lines confirmed the T-DNA-induced IAA biosynthesispathway in the transformed tumor cells. Supplementing auxinto the mutant lines resulted in complete suppression of theshoot-forming ability, but no changes in the endogenous IAAlevels. There was no marked difference in the cytokinin level betweenthe nontransformed callus line and the wild type tumor line.The two mutant lines, however, showed a 20- to 30-fold highercytokinin level which was not affected by the addition of NAA.The T-DNA encoded hormone biosynthetic pathways are discussedin relation to pathways of the host plant. (Received July 29, 1986; Accepted February 14, 1987)     

Effects of visual conditions and prey density on feeding kinetics of paralarvae of Octopus vulgaris from a laboratory spawning

Consumption rate and feeding success of newly hatched paralarvaeof the cephalopod Octopus vulgaris preying on Artemia larvaewere investigated in relation to visual conditions and preydensity. Each paralarva was tested individually using a small-scaleexperimental setup; consumption over one day was measured at20°C. A factorial experiment was designed to investigatethe effects on the consumption rate of two predictor variables:illumination/background (three levels: 7.5 Wm^–2 whitelight + white background, 7.5 Wm^–2 white light +blue background, darkness) and prey density (four levels: 2.35,4.70, 9.40 and 14.10 Artemia metanauplii ml^–1). Consumptionrate varied significantly between different conditions of illuminationand prey density. Light enhanced consumption rate, but differentbackgrounds yielded similar rates. The maximal consumption rateunder illumination was close to 16 Artemia paralarva^–1day^–1, and it was around 5 Artemia paralarva^–1 day^–1for assays in darkness. The predatory efficiency, measured asthe proportion of prey consumed, was significantly affectedby prey density, pointing to a type III functional response.The number of nonfeeding paralarvae was significantly higherin darkness and at low prey density. (Received 14 February 2006; accepted 23 December 2006)     

Down-regulation of trypsinogen expression is associated with growth retardation in alpha1,6-fucosyltransferase-deficient mice: attenuation of proteinase-activated receptor 2 activity

Alpha1,6-fucosyltransferase (Fut8) plays important roles inphysiological and pathological conditions. Fut8-deficient (Fut8^–/–)mice exhibit growth retardation, earlier postnatal death, andemphysema-like phenotype. To investigate the underlying molecularmechanism by which growth retardation occurs, we examined themRNA expression levels of Fut8^–/– embryos (18.5days postcoitum [dpc]) using a cDNA microarray. The DNA microarrayand real-time polymerase chain reaction (PCR) analysis showedthat a group of genes, including trypsinogens 4, 7, 8, 11, 16,and 20, were down-regulated in Fut8^–/– embryos.Consistently, the expression of trypsinogen proteins was foundto be lower in Fut8^–/– mice in the duodenum, smallintestine, and pancreas. Trypsin, an active form of trypsinogen,regulates cell growth through a G-protein-coupled receptor,the proteinase-activated receptor 2 (PAR-2). In a cell culturesystem, a Fut8 knockdown mouse pancreatic acinar cell carcinoma,TGP49-Fut8-KDs, showed decreased growth rate, similar to thatseen in Fut8^–/– mice, and the decreased growth ratewas rescued by the application of the PAR-2-activating peptide(SLIGRL-NH₂). Moreover, epidermal growth factor (EGF)-inducedreceptor phosphorylation was attenuated in TGP49-Fut8-KDs, whichwas highly associated with a reduction of trypsinogens mRNAlevels. The addition of exogenous EGF recovered c-fos, c-jun,and trypsinogen mRNA expression in TGP49-Fut8-KDs. Again, theEGF-induced up-regulation of c-fos and c-jun mRNA expressionwas significantly blocked by the protein kinase C (PKC) inhibitor.Our findings clearly demonstrate a relationship between Fut8and the regulation of EGF receptor (EGFR)-trypsin-PAR-2 pathwayin controlling cell growth and that the EGFR-trypsin-PAR-2 pathwayis suppressed in TGP49-Fut8-KDs as well as in Fut8^–/–mice.     

Roles of complex gangliosides in the development of experimental autoimmune encephalomyelitis

We induced experimental autoimmune encephalomyelitis (EAE) inGM2/GD2 synthase knockout mice (GM2/GD2^–/–), whichcannot synthesize complex gangliosides, such as GM1, GD1a, GD1b,GT1b, and GQ1b, to investigate the roles of complex gangliosidesin the pathogenesis of this disease. We used myelin-oligodendrocyteglycoprotein (MOG) as an immunogen. In active immunization EAE,the severity of clinical score was not different but the diseaseonset was significantly delayed in GM2/GD2^–/– comparedwith those in wild-type mice. When we transferred MOG-reactiveT cells from GM2/GD2^–/– or wild-type mice to wild-typemice, no significant differences were observed between the twogroups. In contrast, when we transferred MOG-reactive T cellsfrom wild-type mice to GM2/GD2^–/– or to wild-typemice, the onset of EAE in GM2/GD2^–/– mice was delayed.The recall response of MOG-specific T cells, the function ofantigen presenting cells, or the expression of several adhesionmolecules in the endothelium were not significantly differentbetween GM2/GD2^–/– and wild-type mice. On the otherhand, quantitative analysis of cellular infiltration in thecentral nervous system (CNS) on day 9 of active immunizationEAE showed that the CD4⁺ cell number in the CNS isolated fromGM2/GD2^–/– mice was significantly less than thatfrom wild-type mice. It indicated that the delayed onset ofEAE in GM2/GD2^–/– mice was due to the delay of themigration of pathogenic T cells into the CNS. Thus, the complexgangliosides may be involved in the T cell–endothelialcell interaction in the pathogenetic process of EAE.     

Beta-adrenergic stimulation does not activate Na+/Ca2+ exchange current in guinea pig, mouse, and rat ventricular myocytes

The effect of -adrenergic stimulation on cardiac Na⁺/Ca²⁺ exchange has been controversial. To clarify the effect, we measured Na⁺/Ca²⁺ exchange current (I_NCX) in voltage-clamped guinea pig, mouse, and rat ventricular cells. When I_NCX was defined as a 5 mM Ni²⁺-sensitive current in guinea pig ventricular myocytes, 1 µM isoproterenol apparently augmented I_NCX by 32%. However, this increase was probably due to contamination of the cAMP-dependent Cl^– current (CFTR-Cl^– current, I_CFTR-Cl), because Ni²⁺ inhibited the activation of I_CFTR-Cl by 1 µM isoproterenol with a half-maximum concentration of 0.5 mM under conditions where I_NCX was suppressed. Five or ten millimolar Ni²⁺ did not inhibit I_CFTR-Cl activated by 10 µM forskolin, an activator of adenylate cyclase, suggesting that Ni²⁺ acted upstream of adenylate cyclase in the -adrenergic signaling pathway. Furthermore, in a low-extracellular Cl^– bath solution, 1 µM isoproterenol did not significantly alter the amplitude of Ni²⁺-sensitive I_NCX at +50 mV, which is close to the reversal potential of I_CFTR-Cl. No change in I_NCX amplitude was induced by 10 µM forskolin. When I_NCX was activated by extracellular Ca²⁺, it was not significantly affected by 1 µM isoproterenol in guinea pig, mouse, or rat ventricular cells. We concluded that -adrenergic stimulation does not have significant effects on I_NCX in guinea pig, mouse, or rat ventricular myocytes. cystic fibrosis transmembrane conductance regulator; nickel ion     

Breast cancer resistance protein (Bcrp1/Abcg2) is expressed in the harderian gland and mediates transport of conjugated protoporphyrin IX

Proper regulation of intracellular levels of protoporphyrin IX (PPIX), the direct precursor of heme, is important for cell survival. A deficiency in ferrochelatase, which mediates the final step in heme biosynthesis, leads to erythropoietic protoporphyria (EPP), a photosensitivity syndrome caused by the accumulation of PPIX in the skin. We have previously shown that mice with a deficiency in the ABC transporter Bcrp1/Abcg2 display a novel type of protoporphyria. This protoporphyria is mild compared with ferrochelatase-dependent EPP, and in itself not sufficient to cause phototoxicity, but it might exacerbate the consequences of other porphyrias. In this study, we identified the mouse harderian gland as a novel expression site of Bcrp1. Because of its pronounced role in porphyrin secretion, the harderian gland presents a useful tool to study the mechanism of Bcrp1-related protoporphyria and transport of porphyrins. Bcrp1^–/– harderian gland displayed a highly increased accumulation of PPIX glycoconjugates, and a similar shift was seen in Bcrp1^–/– liver. Tear- and hepatobiliary excretion data suggest that Bcrp1 controls intracellular levels of PPIX by mediating high affinity transport of its glycoconjugates and possibly low-affinity transport of unconjugated PPIX. This mechanism may allow cells to prevent or reduce cytotoxicity of PPIX under excess conditions, without spillage under physiological conditions where PPIX is needed. ATP binding cassette transporter; knockout mice; protoporphyria     

Isolation, Culture, and Plant Regeneration of Protoplasts of Two Shrubby Oxalis Species from South America

Protoplasts were successfully isolated from internodal callustissues of both Oxalis glaucifolia and O. rhombeo-ovata whenthey were digested in a solution containing 0.1% (w/v) MacerozymeR-10, 0.5% (w/v) cellulase Onozuka R-10 and 0.3 mmol m^–3sucrose. Protoplasts proliferated to give cell colonies on Gamborget al.'s B5 medium supplemented with 0.3 mmol m^–3 mannitol,0.5 mg dm^–32, 4-D, and 2.0 mg dm^–3 kinetin. Calluswas produced upon transfer of cell colonies to Murashige andSkoog medium containing 2.0 mg dm^–3 l-naphthaleneaceticacid (NAA) and 0.1 mg dm^–3 kinetin for O. glaucifolia,or with 5.0 mg dm^–3 NAA and 0.5 mg dm^–3 6-benzylaminopurine,for O. rhombeo-ovata. Plants were regenerated from O. glaucifoliaprotoplasts on a medium containing 0.1 mg dm–3 NAA, 1.0mg dm^–3 kinetin and 1.0 mg dm^–3 gibberellic acid,but only vascular nodules were differentiated by O. rhombeo-ovataprotoplast-derived calli. Key words: Tissue culture, protoplasts, plant regeneration, Oxalis spp     

The alpha2-adrenergic receptor agonist UK 14,304 inhibits secretin-stimulated ductal secretion by downregulation of the cAMP system in bile duct-ligated rats

Secretin stimulates ductal secretion by activation of cAMP PKA CFTR Cl^–/HCO₃^– exchanger in cholangiocytes. We evaluated the expression of _2A-, _2B-, and _2C-adrenergic receptors in cholangiocytes and the effects of the selective ₂-adrenergic agonist UK 14,304, on basal and secretin-stimulated ductal secretion. In normal rats, we evaluated the effect of UK 14,304 on bile and bicarbonate secretion. In bile duct-ligated (BDL) rats, we evaluated the effect of UK 14,304 on basal and secretin-stimulated 1) bile and bicarbonate secretion; 2) duct secretion in intrahepatic bile duct units (IBDU) in the absence or presence of 5-(N-ethyl-N-isopropyl)amiloride (EIPA), an inhibitor of the Na⁺/H⁺ exchanger isoform NHE3; and 3) cAMP levels, PKA activity, Cl^– efflux, and Cl^–/HCO₃^– exchanger activity in purified cholangiocytes. ₂-Adrenergic receptors were expressed by all cholangiocytes in normal and BDL liver sections. UK 14,304 did not change bile and bicarbonate secretion of normal rats. In BDL rats, UK 14,304 inhibited secretin-stimulated 1) bile and bicarbonate secretion, 2) expansion of IBDU luminal spaces, and 3) cAMP levels, PKA activity, Cl^– efflux, and Cl^–/HCO₃^– exchanger activity in cholangiocytes. There was decreased lumen size after removal of secretin in IBDU pretreated with UK 14,304. In IBDU pretreated with EIPA, there was no significant decrease in luminal space after removal of secretin in either the absence or presence of UK 14,304. The inhibitory effect of UK 14,304 on ductal secretion is not mediated by the apical cholangiocyte NHE3. ₂-Adrenergic receptors play a role in counterregulating enhanced ductal secretion associated with cholangiocyte proliferation in chronic cholestatic liver diseases. bicarbonate secretion; chloride efflux; gastrointestinal hormones; intrahepatic biliary epithelium; protein kinase A     

Comparative mapping of HKT genes in wheat, barley, and rice, key determinants of Na+ transport, and salt tolerance

Salt tolerance of plants depends on HKT transporters (High-affinityK⁺ Transporter), which mediate Na⁺-specific transport or Na⁺-K⁺co-transport. Gene sequences closely related to rice HKT geneswere isolated from hexaploid bread wheat (Triticum aestivum)or barley (Hordeum vulgare) for genomic DNA southern hybridizationanalysis. HKT gene sequences were mapped on chromosomal armsof wheat and barley using wheat chromosome substitution linesand barley–wheat chromosome addition lines. In addition,HKT gene members in the wild diploid wheat ancestors, T. monococcum(A^m genome), T. urartu (A^u genome), and Ae. tauschii (D^t genome)were investigated. Variation in copy number for individual HKTgene members was observed between the barley, wheat, and ricegenomes, and between the different wheat genomes. HKT2;1/2-like,HKT2;3/4-like, HKT1;1/2-like, HKT1;3-like, HKT1;4-like, andHKT1;5-like genes were mapped to the wheat–barley chromosomegroups 7, 7, 2, 6, 2, and 4, respectively. Chromosomal regionscontaining HKT genes were syntenic between wheat and rice exceptfor the chromosome regions containing the HKT1;5-like gene.Potential roles of HKT genes in Na⁺ transport in rice, wheat,and barley are discussed. Determination of the chromosome locationsof HKT genes provides a framework for future physiological andgenetic studies investigating the relationships between HKTgenes and salt tolerance in wheat and barley. Key words: Barley, comparative mapping, HKT, rice, salt tolerance, sodium transport, wheat     

Respiration rates of dominant hydromedusae in the North Inlet tidal estuary during winter and summer

The objective of this study was to quantify the respiratoryrequirements and oxygen consumption rates for seasonal hydromedusaepopulations in the North Inlet estuary, Georgetown, SC, USA.Respiration rates were determined for hydromedusae collectedin January 2006, 2007 and July 2006 using microrespirometry.The numerically dominant gelatinous predators were Nemopsisbachei (January 2006 and 2007) and Bougainvillia muscus (July2006). Seasonal oxygen consumption rates were low (0.80–1.27µmol O₂ g^–1 h^–1). Q₁₀ values were high duringJanuary 2006 and July 2006 (4.7), but low during January 2007(0.5), suggesting hydromedusae were sensitive to the exposuretemperatures (7–34°C) in this study and well adaptedto the temperature range encountered in this estuary. Moreover,the oxygen consumption rates during July 2006 showed significantdifferences between the two species. The allometric relationshipbetween respiration rate and hydromedusae weight differed seasonally.Our findings suggest hydromedusae can persist in hypoxic conditionsand may partially explain the frequently observed increase inthese cnidarians under conditions of low dissolved oxygen concentrations.     

Population biomass, feeding, respiration and growth rates, and carbon budget of the scyphomedusa Aurelia aurita in the Inland Sea of Japan

We investigated the seasonal occurrence, wet : dry : carbon: nitrogen weight ratios, population biomass, gastric pouchcontents, and rates of feeding, growth and respiration of thescyphomedusa Aurelia aurita in the central part of the InlandSea of Japan. Aurelia aurita medusae began to appear in January/Februaryas ephyrae, reached annual maximum body size in July/August,and disappeared, presumably due to death, by November. Initialslow growth in early spring was followed by a period of exponentialgrowth (mean growth rate: 0.069 d^–1) between April andJuly. In the Ondo Strait, which is characterized by strong tidalmixing, the A. aurita population (mean carbon biomass: 66.0mg C m^–3) overwhelmingly dominated the zooplankton-communitybiomass (mean biomass of micro- and mesozooplankton: 23.7 mgC m^–3) between May and early August The gastric contentanalysis revealed that A. aurita ate almost all micro- and mesozooplankters,of which small copepods were most important. On the basis ofdigestion time for small copepods (60 min) and their abundancein the gastric pouch of field-collected A. aurita, we determinedthe weight specific feeding rates and clearance rates. The formerincreases linearly with increasing copepod abundance, but thelatter was relatively constant irrespective of the food supply.We also measured the respiration rates of A. aurita and expressedthem as functions of body weight and temperature. These physio-ecologicalparameters enabled us to construct the carbon budget of theA. aurita population typical of early summer in the Ondo Strait.Predicted population-feeding rate (6.07 mg C m^–3 d^–1)was higher than the population-food requirement for both metabolismand growth (4.55 mg C m^–3 d^–1), indicating thatfood supply was sufficient to sustain the observed growth rate.This feeding rate was equivalent to 26% of micro- and mesozooplanktonbiomass, a significant impact on zooplankton.     

Different Intracellular Localization of Two Cytochrome P-450 Systems, Ipomeamarone 15-Hydroxylase and Cinnamic Acid 4-Hydroxylase, in Sweet Potato Root Tissue Infected with Ceratocystis fimbriata

Ipomeamarone 15-hydroxylase activity was mainly recovered inthe pellet fraction between centrifugations at 10,000 and 100,000?gfrom a crude extract of Ceratocystis fimbriata-infected sweetpotato root tissue, whereas cinnamic acid 4-hydroxylase activitywas found between centrifugations at 300 and 10,000?g. Whenparticles in the crude extract were fractionated by sucrosedensity gradient centrifugation, the rough-surfaced microsomeswere distributed over a wide density range from 1.09 to 1.14g cm^–3, judging from the distributions of protein, RNAand NADPH-cytochrome c reductase activity. Phosphorylcholine-glyceridetransferase activity was only in the lighter half of the microsomalfraction (density: 1.09–1.11 g cm^–3). Ipomeamarone15-hydroxylase activity was found in heavier half of the microsomalfraction (density: 1.10–1.14 g cm^–3). We proposethat this tissue has two rough-surfaced endoplasmic reticulumspecies, only one of which carries phosphorylcholine-glyceridetransferase, and that the cytochrome P-450 system is localizedon the species lacking the enzyme. Cinnamic acid 4-hydroxylaseactivity was mainly found in a fraction that had densities of1.17–1.19 g cm^–3 and contained vesicular particlesof various sizes. ¹ Present address: Laboratory of Food Hygienics, Faculty ofAgriculture, Kagawa University, Miki-cho, Kida-gun, Kagawa 761-07,Japan. (Received September 6, 1984; Accepted December 27, 1984)     

Measurement of Yield Threshold and Cell Wal Extensibility of Intact Wheat Roots under Different Ionic, Osmotic and Temperature Treatments

Yield stress threshold (Y) and volumetric extensibility () arethe rheological properties that appear to control root growth.In this study they were measured in wheat roots by means ofparallel measurement of the growth rate (r) of intact wheatroots and of the turgor pressures (P) of individual cells withinthe expansion zone. Growth and turgor pressure were manipulatedby immersion in graded osmoticum (mannitol) solutions. Turgorwas measured with a pressure probe and growth rate by visualobservation. The influence of various growth conditions on Yand was investigated; (a) At 27 °C.In 0.5 mol m^–3 CaCl₂ r, P, Y and were20.7±4.6 µm min^–1, 0.77±0.05 MPa,0.07±0.03 MPa and 26±1.9 µm min^–1MPa^–1 (expressed as increase in length), respectively.Following 24 h growth in 10 mol m^–3 KC1 these parametersbecame 12.3±3.5 µm min^–1, 0.72±0.04MPa, 0.13±0.01 MPa and 21±0.7 µm min^–1MPa^–1. After 24 h osmotic adjustment in 150 mol m^–3mannitol/0.5 mol m^–3 CaCl₂ r= 19.6±4.2 µmmin^–1, P = 0.68±0.05 MPa and Y and were 0.07±0.04MPa and 30±0.2 µm min^–1 MPa^–01, respectively.After 24 h growth in 350 mol m^–3 mannitol/0.5 mol m^–3CaCl₂ r= 13.3±4.1 µm min^–1, P= 0.58±0.07MPa, Y=0.12±0.01 MPa and ø 32±0.2 tim min^–1MPa^–1. During osmotic adjustment in 200 mol m^–3mannitol/0.5 mol m^–3 CaCl₂, with or without KCl, the recoveryof growth rate corresponded to turgor pressure recovery (t1/2approximately 3 h). (b) At 15 °C. Lowered temperature dramatically influencedthe growth parameters which became r= 8.3±2.8 um min^–1,P=0.78 MPa, r=<0.2 MPa and =15±0.1 µm min^–1MPa^–1. Therefore, Y and are influenced by 10 mol m^–3 K⁺ ionsand low temperature. In each case the effective pressure forgrowth (P-Y) was large indicating that small fluctuations ofsoil water potential will not stop root elongation. Key words: Yield threshold, cell wall extensibility, wheat root growth, temperature, turgor pressur     

Structure, function, and genomic organization of human Na(+)-dependent high-affinity dicarboxylate transporter

We have clonedand functionally characterized the human Na⁺-dependenthigh-affinity dicarboxylate transporter (hNaDC3) from placenta. ThehNaDC3 cDNA codes for a protein of 602 amino acids with 12 transmembrane domains. When expressed in mammalian cells, the clonedtransporter mediates the transport of succinate in the presence ofNa⁺ [concentration of substrate necessary for half-maximaltransport (K_t) for succinate = 20 ± 1 µM]. Dimethylsuccinate also interacts with hNaDC3. TheNa⁺-to-succinate stoichiometry is 3:1 and concentration ofNa⁺ necessary for half-maximal transport(K^Na+_0.5) is 49 ± 1 mM as determined by uptake studies withradiolabeled succinate. When expressed in Xenopuslaevis oocytes, hNaDC3 induces Na⁺-dependent inwardcurrents in the presence of succinate and dimethylsuccinate. At amembrane potential of 50 mV,K^Suc_0.5 is 102 ± 20 µM andK^Na+_0.5 is 22 ± 4 mM as determined by the electrophysiological approach. Simultaneous measurements of succinate-evoked charge transfer andradiolabeled succinate uptake in hNaDC3-expressing oocytes indicate acharge-to-succinate ratio of 1:1 for the transport process, suggestinga Na⁺-to-succinate stoichiometry of 3:1. pH titration ofcitrate-induced currents shows that hNaDC3 accepts preferentially thedivalent anionic form of citrate as a substrate. Li⁺inhibits succinate-induced currents in the presence of Na⁺.Functional analysis of rat-human and human-rat NaDC3 chimeric transporters indicates that the catalytic domain of the transporter lies in the carboxy-terminal half of the protein. The humanNaDC3 gene is located on chromosome20q12-13.1, as evidenced by fluorescent in situ hybridization. Thegene is >80 kbp long and consists of 13 exons and 12 introns.

     

'Nonparametric inference in multivariate mixtures' * Biometrika (2005), 92, pp. 667 678

The left-hand side of equation (2·8), on p. 671, shouldread {₁ (1 – ₁)}^–1/2 (2₁ – 1) rather than{(1 – ₁)/₁}^1/2 (2₁ – 1). Reflecting this change,the left-hand side of equation (3·1) on the same pageshould be altered to , and the formula at the foot of p. 677 should be modified to {₁ (1– ₁)}^–1/2 (2₁ – 1) + O_p(n^–1/2). No otherformula is affected, and the left-hand side of (2·8)is still increasing in ₁. The numerical results, discussed in4, are influenced in minor ways. In the simulation study, absolutebias is reduced, and variance is either slightly increased orslightly decreased. In the real-data example, using the nonparametricapproach to analysis, mean squared error is further reduced,from 0·0011 to 0·0004. We are grateful to HiroKasahara and Katsumi Shimotsu for pointing out the error.     

Growth and development of Neocalanus flemingeri/plumchrus in the northern Gulf of Alaska: validation of the artificial-cohort method in cold waters

In situ growth and development of Neocalanus flemingeri/plumchrusstage C1–C4 copepodites were estimated by both the artificial-cohortand the single-stage incubation methods in March, April andMay of 2001–2005 at 5–6°C. Results from thesetwo methods were comparable and consistent. In the field, C1–C4stage durations ranged from 7 to >100 days, dependent ontemperature and chlorophyll a (Chl a) concentration. Averagestage durations were 12.4–14.1 days, yielding an averageof 56 days to reach C5, but under optimal conditions stage durationswere closer to 10 days, shortening the time to reach C5 (fromC1) to 46 days. Generally, growth rates decreased with increasingstage, ranging from 0.28 day^–1 to close to zero but weretypically between 0.20 and 0.05 day^–1, averaging 0.110± 0.006 day^–1 (mean ± SE) for single-stageand 0.107 ± 0.005 day^–1 (mean ± SE) forartificial-cohort methods. Growth was well described by equationsof Michaelis–Menten form, with maximum growth rates (G_max)of 0.17–0.18 day^–1 and half saturation Chl a concentrations(K_chl) of 0.45–0.46 mg m^–3 for combined C1–3,while G_max dropped to 0.08–0.09 day^–1 but K_chl remainedat 0.38–0.93 mg m^–3 for C4. In this study, in situgrowth of N. flemingeri/plumchrus was frequently food limitedto some degree, particularly during March. A comparison withglobal models of copepod growth rates suggests that these modelsstill require considerable refinement. We suggest that the artificial-cohortmethod is the most practical approach to generating the multispeciesdata required to address these deficiencies.     

Effects of Colchicine, Vinblastine, Cytochalasin B and Phalloidin on the Seismonastic Movement of Mimosa pudica Leaf and on Motor Cell Ultrastructure

Fleurat-Lessard, P., Roblin, G., Bonmort, J. and Besse, C. 1988.Effects of colchicine, vinblastine, cytochalasin B and phalloidinon the seismonastic movement of Mimosa pudica leaf and on motorcell ultrastructure.—J. exp. Bot. 39: 209–221. Colchicine at 1 x 10^–3 mol dm^–3 does not affectthe seismonastic movement of Mimosa pudica leaves but disruptsmicrotubules in motor cells. Vinblastine at 5 x 10^–3 moldm^–3 does not affect this movement and partly disruptsmicrotubules. Vinblastine at 1 x 10^–4 mol dm^–3 alwaysdisrupts microtubules, even after a 12 h reversibility whenthe movement is restored. These drugs, applied at the same respectiveconcentrations, do not alter cytoplasmic and vacuolar fibrils.Cytochalasin B and phalloidin alter the seismonastic movementof Mimosa leaves when applied at concentrations of 1.25 x 10^–3and 2.4 x 10^–4 mol dm^–3 respectively. These drugs,used at the same respective concentrations, also affect themotor cell structure and, in particular, modify the arrangementand the structure of the fibrils but they do not destroy themicrotubules. These data suggest that microtubules are not directly involvedin the seismonastic reaction whereas fibrils, formed by thin(3.0 nm wide) filaments, may be implicated in this reaction. Key words: Colchicine, cytochalasin B, phalloidin, Mimosa pudica, motor cells, vinblastine