Kinetics of NO3- Influx in Spruce

Influxes of 13NO3- across the root plasmalemma were measured in intact seedlings of Picea glauca (Moench) Voss. Three kinetically distinct uptake systems for NO3- were identified. In seedlings not previously exposed to external NO3-, a single Michaelis-Menten-type constitutive high-affinity transport system (CHATS) operated in a 2.5 to 500 [mu]M range of external NO3- [NO3-]o. The Vmax of this system was 0.1 [mu]mol g-1 h-1, and the Km was approximately 15 [mu]M. Following exposure to NO3- for 3 d, this CHATS activity was increased approximately 3-fold, with no change of Km. In addition, a NO3--inducible high-affinity system became apparent with a Km of approximately 100[mu]M. The combined Vmax for these discrete saturable components was 0.7 [mu]mol g-1 h-1. In both uninduced and induced plants a linear low-affinity system, additive to CHATS and an NO3--inducible high-affinity system, operated at [NO3-]o [greater than or equal to] 1 mM. The time taken to achieve maximal rates of uptake (full induction) was 2 d from 1.5 mM [NO3-]o and 3 d from 200 [mu]M [NO3-]o.     

In vivo cloning of a carboxy-terminalrpoB allele which confers altered transcriptional properties

A 3′-terminal mutation of the gene encoding the β subunit ofEscherichia coli RNA polymerase was isolated using anin vivo polA(Ts) technique. Cloning of the allele was monitored by virtue of the fact that the deletion Δ(rpoB) 1570-1 resulted in an altered-size restriction fragment. DNA sequencing confirmed the predicted nature and location of the mutation: Δ(rpoB) 1570-1 involved an in-frame deletion of 186 bp (62 codons) encoding amino acid residues 967–1028. The phenotype conferred by Δ(rpoB) 1570-1 is discussed with respect to conserved domains within the β polypeptide. Dedicated to Dr. J. Spížek on the occasion of his 60th birthday     

Transfection,expression, and DNA sequence of a gene encoding a BoLA-A11 antigen

The nucleotide sequence data reported in this paper have been submitted to the EMBL, GenBank, and DDBJ nucleotide sequence databases, and have been assigned the accession numbers X82671–X82675     

Characterization of the NADH dehydrogenase and fumarate reductase of Fibrobacter succinogenes subsp. succinogenes S85

Conditions promoting maximal in vitro activity of the particulate NADH:fumarate reductase from Fibrobacter succinogenes were determined. This system showed a pH optimum of 6.0 in K⁺ MES buffer only when salt (NaCl or KCl) was present. Salt stimulated the activity eightfold at the optimal concentration of 150m M. This effect was due to stimulation of fumarate reductase activity as salt had little effect on NADH: decylubiquinone oxidoreductase (NADH dehydrogenase). The stimulation of fumarate reductase by salt at pH 6.0 was not due to removal of oxaloacetate from the enzyme. Kinetic parameters for several inhibitors were also measured. NADH dehydrogenase was inhibited by rotenone at a single site with a K _i of 1 M. 2-Heptyl-4-hydroxyquinonline-N-oxide (HOQNO) inhibited NADH: fumarate reductase with a K _i of 0.006 M, but NADH dehydrogenase exhibited two HOQNO inhibition constants of approximately 1 M and 24 M. Capsaicin and laurylgallate each inhibited NADH dehydrogenase by only 20% at 100 M. NADH dehydrogenase gave K _m values of 1 M for NADH and 4 M for reduced hypoxanthine adenine dinucleotide.Published with the approval of the Director of the Agricultural Experiment Station, North Dakota State University, as journal article no. 2201     

Perception of effort during high-intensity exercise at low, moderate and high wet bulb globe temperatures

The purpose of this study was to determine the effect of low, moderate and high wet bulb globe temperatures (T _wbg) on cardiovascular variables and ratings of perceived exertion (RPE) during moderately prolonged, high-intensity exercise. Six subjects [four men and two women; mean (SD) age, 22.0 (1.2) years; maximum oxygen consumption ({ie519-1}), 51.0 (8.4) ml · kg^–1 · min^–1] completed 30 min of exercise (80% {ie519-2}) on a cycle ergometer at low [14.7 (2.1)°C], moderate [21.0 (1.5)° C], and high [27.4 (2.3)° C]T _wbg. Two additional subjects completed 20 min of exercise in the high temperature condition, but completed 30 min in the moderate and lowT _wbg. Heart rate (f _c), blood pressure, blood lactate (La), mean skin temperature ( _sk), , and RPE were measured at 10, 20 and 30 min. Results showed thatf _c, rate pressure product, RPE, pulmonary ventilation and ventilatory equivalent for oxygen increased (P < 0.05) across time for all conditions, while decreased across time. _sk andf _c were significantly greater across time in the high condition [35.9 (0.65)° C; 176 (12.6) beats · min^–1] compared to the moderate [34.6 (1.5)° C; 170 (17.2) beats · min^–1] and the low condition [31.7 (1.5)° C; 164 (17.1) beats-min^–1]. However, there were no differences throughout exercise in RPE [high,.16.2 (2.0); moderate, 16.4 (2.2); low, 16.3 (1.9)] and across the conditions. These data suggest that RPE is closely related to metabolic intensity but is not a valid indicator of cardiovascular strain during exercise in highT _wbg conditions.     

The effects of temperature and hyperoxia on arterial PO2 and acid-base status in Piaractus mesopotamicus

The effects of hyperoxia and change of temperature (range 20–30° C) on blood gases were studied in the teleost fish Piaractus mesopotamicus , native to several major river systems in Brazil. Large hyperoxia-induced increases of arterial P o₂ ( P _ao₂) indicated that true branchial blood shunts are negligible in relation to total gill perfusion. This implies that blood gases will be influenced by ventilation rather than by shunts. Acute variations of temperature ( t ) were accompanied by changes of arterial blood pH (on the average Δ p H_aΔt⁻¹ of — 0·015 units °C⁻¹), due mainly to alterations of P _aco₂: 2·4 mmHg at 20° C, 5·0 mmHg at 30° C. Concomitantly, P _ao₂ declined from 116 mmHg (20° C) to 89 mmHg (30° C). The data suggest that temperature-induced changes of acid-base status depend mainly on gill ventilation and that the decrease of P _ao₂ with higher temperature is a result of this regulation.     

Bioethics for clinicians: 7. Truth telling

The standard of professional candour with patients has undergone a significant change over the past 30 years. Independent of their obligation to disclose information necessary for informed consent, physicians are increasingly expected to communicate important information to patients that is not immediately related to treatment decisions. The purpose of truth telling is not simply to enable patients to make informed choices about health care and other aspects of their lives but also to inform them about their situation. Truth telling fosters trust in the medical profession and rests on the respect owed to patients as persons. It also prevents harm, as patients who are uninformed about their situation may fail to get medical help when they should.     

Studies of the Uptake of Nitrate in Barley : II. Energetics

Q₁₀ values for ¹³NO₃⁻ influx were determined in `uninduced' (NO<sub>3</sub><sup>−</sup>-starved) and `induced' (NO₃⁻-pretreated) roots of barley (Hordeum vulgare L.) plants at various concentrations of external NO₃⁻ ([NO₃⁻]₀). At 0.02 mole per cubic meter [NO₃⁻]₀, Q₁₀ values for influx were from 3 to 4 between 5 and 10°C. As [NO₃⁻]₀ increased Q₁₀ values decreased, reaching values of 1.2 and 2.0, respectively, at 20 moles per cubic meter in uninduced and induced plants. The metabolic dependence of ¹³NO₃⁻ influx at low and high [NO₃⁻]₀ (0.1 and 20.0 moles per cubic meter, respectively) in uninduced and induced plants was probed by the use of various inhibitors. These experiments confirmed the findings of the Q₁₀ studies, demonstrating that at low [NO₃⁻]₀¹³NO₃⁻ influx was extremely sensitive to metabolic inhibition. By contrast, at high [NO₃⁻]₀, influx was relatively insensitive to the presence of inhibitors.     

Mobilization of iron from endocytic vesicles. The effects of acidification and reduction

The factors necessary to dissociate iron from transferrin in endocytic vesicles and to mobilize the iron across the vesicle membrane were studied in a preparation of endocytic vesicles markedly enriched in transferrin-transferrin receptor complexes isolated from rabbit reticulocytes. Vesicles were prepared with essentially fully saturated transferrin by incubating the reticulocytes with the protonophore carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone prior to incubation with 59Fe, 125I-transferrin with or without fluorescein isothiocyanate labeling. Initiation of acidification by the addition of ATP was sufficient to achieve dissociation of 59Fe from transferrin with a rate constant of 0.054 +/- 0.06 s-1. Mobilization of 59Fe out of the vesicles required, besides ATP, the addition of a reductant with 1 mM ascorbate, allowing approximately 60% mobilization at 10 min with a rate constant of 0.0038 +/- 0.0006 s-1. An NADH:ferricyanide reductase activity could be demonstrated in the vesicles with an activity of 7.1 x 10(-9) mol of NADH reduced per min/mg of vesicle protein. Both dissociation and mobilization were inhibited by N-ethylmaleimide, carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone, and monensin. Mobilization, but not dissociation, was inhibited by the permeant Fe(II) chelator alpha,alpha'-dipyridyl. The Fe(III) chelators deferoxamine, diethylenetriaminepentaacetic acid, and apotransferrin did not promote mobilization of dissociated iron in the absence of a reductant. This study establishes the basis for the cellular incorporation of iron through the endocytic pathway in which the endocytic vesicle membrane utilizes, in a sequential way, an acidification system, an iron reduction system, and an Fe(II) transporter system.