Evaluation of uptake and transport of ultrasmall superparamagnetic iron oxide nanoparticles by human brain-derived endothelial cells.

Aim: Ultrasmall superparamagnetic iron oxide nanoparticles (USPIO-NPs) are under development for imaging and drug delivery; however, their interaction with human blood-brain barrier models is not known. Materials & Methods: The uptake, reactive oxygen species production and transport of USPIO-NPs across human brain-derived endothelial cells as models of the blood-brain tumor barrier were evaluated for either uncoated, oleic acid-coated or polyvinylamine-coated USPIO-NPs. Results: Reactive oxygen species production was observed for oleic acid-coated and polyvinylamine-coated USPIO-NPs. The uptake and intracellular localization of the iron oxide core of the USPIO-NPs was confirmed by transmission electron microscopy. However, while the uptake of these USPIO-NPs by cells was observed, they were neither released by nor transported across these cells even in the presence of an external dynamic magnetic field. Conclusion: USPIO-NP-loaded filopodia were observed to invade the polyester membrane, suggesting that they can be transported by migrating angiogenic brain-derived endothelial cells.

Role of ATP-dependent potassium channels in pulmonary vascular tone of fetal lambs with congenital diaphragmatic hernia.

High mortality in newborn babies with congenital diaphragmatic hernia (CDH) is principally due to persistent pulmonary hypertension. ATP-dependent potassium (K(ATP)) channels might modulate pulmonary vascular tone. We have assessed the effects of Pinacidil, a K(ATP) channel opener, and glibenclamide (GLI), a K(ATP) channel blocker, in near full-term lambs with and without CDH. In vivo, pulmonary hemodynamics were assessed by means of pressure and blood flow catheters. In vitro, we used isolated pulmonary vessels and immunohistochemistry to detect the presence of K(ATP) channels in pulmonary tissue. In vivo, pinacidil (2 mg) significantly reduced pulmonary vascular resistance (PVR) in both controls and CDH animals. GLI (30 mg) significantly increased pulmonary arterial pressure (PAP) and PVR in control animals only. In vitro, pinacidil (10 microM) relaxed, precontracted arteries from lambs with and without CDH. GLI (10(-5) microM) did not raise the basal tone of vessels. We conclude that activation of K(ATP) channels could be of interest to reduce pulmonary vascular tone in fetal lambs with CDH, a condition often associated with persistent pulmonary hypertension of the newborn.

Faster oxygen uptake kinetics during recovery is related to better repeated sprinting ability.

This study was designed to test the hypothesis that subjects having faster oxygen uptake (VO(2)) kinetics during off-transients to exercises of severe intensity would obtain the smallest decrement score during a repeated sprint test. Twelve male soccer players completed a graded test, two severe-intensity exercises, followed by 6 min of passive recovery, and a repeated sprint test, consisting of seven 30-m sprints alternating with 20 s of active recovery. The relative decrease in score during the repeated sprint test was positively correlated with time constants of the primary phase for the VO(2) off-kinetics (r = 0.85; p < 0.001) and negatively correlated with the VO(2) peak (r = -0.83; p < 0.001). These results strengthen the link found between VO(2) kinetics and the ability to maintain sprint performance during repeated sprints.

Selectivity in the binding of psychotropic drugs to the variants of alpha-1 acid glycoprotein.

The S- and F-forms of alpha-1 acid glycoprotein (AAG) variants have been isolated by isoelectric focusing with immobilines from commercially available AAG. In equilibrium dialysis experiments using a multicompartmental system, a higher affinity for various basic drugs has been found with S- in comparison with F-AAG: Amitriptyline, nortriptyline, imipramine, desipramine, trimipramine, methadone, thioridazine, clomipramine, desmethylclomipramine, and maprotiline. The selectivity (binding to S- vs. F-AAG) is the most pronounced for methadone and the lowest for thioridazine, while it is absent for the acidic drug mephenytoin.

Effects of sodium sulphate and potassium chloride fertilizers on the nutritive value of timothy grown on different soils

Selostus: Natrium- ja kaliumlannoituksen vaikutus timotein ravintoarvoon

Glucose uptake, utilization, and signaling in GLUT2-null islets.

We previously reported that pancreatic islet beta-cells from GLUT2-null mice lost the first phase but preserved the second phase of glucose-stimulated insulin secretion (GSIS). Furthermore, we showed that the remaining secretory activity required glucose uptake and metabolism because it can be blocked by inhibition of oxidative phosphorylation. Here, we extend these previous studies by analyzing, in GLUT2-null islets, glucose transporter isoforms and glucokinase expression and by measuring glucose usage, GSIS, and glucose-stimulated insulin mRNA biosynthesis. We show that in the absence of GLUT2, no compensatory expression of either GLUT1 or GLUT3 is observed and that glucokinase is expressed at normal levels. Glucose usage by isolated islets was increased between 1 and 6 mmol/l glucose but was not further increased between 6 and 20 mmol/l glucose. Parallel GSIS measurements showed that insulin secretion was not stimulated between 2.8 and 6 mmol/l glucose but was increased by >4-fold between 6 and 20 mmol/l glucose. Stimulation by glucose of total protein and insulin biosynthesis was also markedly impaired in the absence of GLUT2. Finally, we re-expressed GLUT2 in GLUT2-null beta-cells using recombinant lentiviruses and demonstrated a restoration of normal GSIS. Together, these data show that in the absence of GLUT2, glucose can still be taken up by beta-cells, albeit at a low rate, and that this transport activity is unlikely to be attributed to GLUT1 or GLUT3. This uptake activity, however, is limiting for normal glucose utilization and signaling to secretion and translation. These data further demonstrate the key role of GLUT2 in murine beta-cells for glucose signaling to insulin secretion and biosynthesis.

Brain lactate kinetics: Modeling evidence for neuronal lactate uptake upon activation.

A critical issue in brain energy metabolism is whether lactate produced within the brain by astrocytes is taken up and metabolized by neurons upon activation. Although there is ample evidence that neurons can efficiently use lactate as an energy substrate, at least in vitro, few experimental data exist to indicate that it is indeed the case in vivo. To address this question, we used a modeling approach to determine which mechanisms are necessary to explain typical brain lactate kinetics observed upon activation. On the basis of a previously validated model that takes into account the compartmentalization of energy metabolism, we developed a mathematical model of brain lactate kinetics, which was applied to published data describing the changes in extracellular lactate levels upon activation. Results show that the initial dip in the extracellular lactate concentration observed at the onset of stimulation can only be satisfactorily explained by a rapid uptake within an intraparenchymal cellular compartment. In contrast, neither blood flow increase, nor extracellular pH variation can be major causes of the lactate initial dip, whereas tissue lactate diffusion only tends to reduce its amplitude. The kinetic properties of monocarboxylate transporter isoforms strongly suggest that neurons represent the most likely compartment for activation-induced lactate uptake and that neuronal lactate utilization occurring early after activation onset is responsible for the initial dip in brain lactate levels observed in both animals and humans.

CDK9 regulates AR promoter selectivity and cell growth through serine 81 phosphorylation.

Previously we determined that S81 is the highest stoichiometric phosphorylation on the androgen receptor (AR) in response to hormone. To explore the role of this phosphorylation on growth, we stably expressed wild-type and S81A mutant AR in LHS and LAPC4 cells. The cells with increased wild-type AR expression grow faster compared with parental cells and S81A mutant-expressing cells, indicating that loss of S81 phosphorylation limits cell growth. To explore how S81 regulates cell growth, we tested whether S81 phosphorylation regulates AR transcriptional activity. LHS cells stably expressing wild-type and S81A mutant AR showed differences in the regulation of endogenous AR target genes, suggesting that S81 phosphorylation regulates promoter selectivity. We next sought to identify the S81 kinase using ion trap mass spectrometry to analyze AR-associated proteins in immunoprecipitates from cells. We observed cyclin-dependent kinase (CDK)9 association with the AR. CDK9 phosphorylates the AR on S81 in vitro. Phosphorylation is specific to S81 because CDK9 did not phosphorylate the AR on other serine phosphorylation sites. Overexpression of CDK9 with its cognate cyclin, Cyclin T, increased S81 phosphorylation levels in cells. Small interfering RNA knockdown of CDK9 protein levels decreased hormone-induced S81 phosphorylation. Additionally, treatment of LNCaP cells with the CDK9 inhibitors, 5,6-dichloro-1-β-D-ribofuranosylbenzimidazole and Flavopiridol, reduced S81 phosphorylation further, suggesting that CDK9 regulates S81 phosphorylation. Pharmacological inhibition of CDK9 also resulted in decreased AR transcription in LNCaP cells. Collectively these results suggest that CDK9 phosphorylation of AR S81 is an important step in regulating AR transcriptional activity and prostate cancer cell growth.

Relationship between potassium fertilization and panicle blast severity in upland rice

The influence of K2O (0, 40, 80, 120 kg ha-1) at varying rates of N application (0, 30, 60 kg ha-1) at planting, on panicle blast (Pyricularia grisea (Cooke) Sacc.) was studied in a field experiment conducted during three consecutive years with the upland rice cultivar Douradão. Panicle blast severity decreased with increasing rates of potassium in the absence of nitrogen (N0). The relationship between panicle blast and K rates was quadratic at 30 kg ha-1 of nitrogen. Significant response to K fertilization was not obtained at 60 kg ha-1 of nitrogen in relation to panicle blast severity.

Effect of phosphorus and potassium on yield and on agronomical characteristics of cotton

With the objective of studying the effect of increasing phosphorus and potassium doses on the agronomical and technological characteristics of the cotton (Gossypium hirsutum L.), cultivar IAC 20, an experiment was carried out during 1994/95 on a Red-Dark Latossol at the Embrapa-Centro de Pesquisa Agropecuária do Oeste (CPAO) in Ponta Porã, MS, Brazil. A randomized bloch design was used in a 3 x 5 factorial arrangement with four replications. The doses were 30, 60 and 90 kg ha-1 of P2O5, applied as triple superphosphate, and 0, 30, 60, 90 and 120 kg ha-1 of K2O as KCl. The K2O doses used had a significant influence on the seed cotton yield, plant height and weight of 100 seeds and of bolls.

Fruit size, mineral composition and quality of trickle-irrigated tomatoes as affected by potassium rates

An experiment was conducted to determine the fruit size, mineral composition and quality of trickle-irrigated tomatoes as affected by potassium fertilizer rates. Six potassium (K) rates were applied as KCl, corresponding to 0, 48.4, 118.6, 188.8, 259.0 and 399.4 kg ha-1, with four replicates, following a randomized block design. Quadratic responses to K rates were observed for double extra large (diameter > 60 mm), extra large (56 to 60 mm) and large (52 to 56 mm) fruit yields. Maximum yields of these classes were achieved with K rates of 116, 190 and 233 kg ha-1, respectively. Fruit dry matter, phosphorus, sulfur and magnesium contents were not affected by K rates, but nitrate and K contents showed significant increments as K rates were increased. Vitamin C, total soluble solids, lycopene and beta-carotene contents in the fruits were not affected by K rates. Increments in the K rate lowered the fruit pH and increased total acids content.

Tomato yield and potassium concentrations in soil and in plant petioles as affected by potassium fertirrigation

Tomato (Lycopersicon esculentum Mill.) cv. Santa Clara was grown on a silt clay soil with 46 mg dm-3 Mehlich 1 extractable K, to evaluate the effects of trickle-applied K rates on fruit yield and to establish K critical concentrations in soil and in plant petioles. Six potassium rates (0, 48, 119, 189, 259 and 400 kg ha-1 K) were applied in a randomized complete block design with four replications. Soil and plant K critical levels were determined at two plant growth stages (at the beginning of the second and fourth cluster flowering). Total, marketable and weighted yields increased with K rates, reaching their maximum of 86.4, 73.4, and 54.9 ton ha-1 at 198, 194, and 125 kg ha-1 K , respectively. At the first soil sampling date K critical concentrations in the soil associated with K rates for maximum marketable and weighted yields were 92 and 68 mg dm-3, respectively. Potassium critical concentrations in the dry matter of the petioles sampled by the beginning of the second and fourth cluster flowering time, associated with maximum weighted yield, were 10.30 and 7.30 dag kg-1, respectively.

Estimating potassium nutrition of Norway spruce with needle analysis during different seasons

Tiivistelmä: Kuusen neulasanalyyttinen kaliumravitsemuksen arviointi eri vuodenaikoina