In vivo effects of the controlled NO donor/scavenger ruthenium cyclam complexes on blood pressure

Ruthenium(II/III) complexes able to bind and release NO* were tested in vivo, in conscious Wistar rats instrumented for continuous blood pressure (BP) measurement and administration of in bolus injections (5 to 100 nmol/Kg i.v.) of trans-[Ru(II)Cl(NO+)(cyclam)](PF6)2 (cyclam-NO) or sodium nitroprusside (SNP). For normotensive rats, cyclam-NO produced a sustained 10% BP reduction of basal MAP during 7 +/- 0.4 to 11 +/- 0.3 min. In acute hypertensive rats, cyclam-NO produced BP reduction 3-fold larger than in normotensive rats and similar to that of SNP (maximal effect: 41 +/- 1.3 vs. 45 +/- 2.2 mmHg, respectively). However, the duration of the effect of cyclam-NO was 13 to 21-fold longer than that of SNP. The hypotensive effect of cyclam-NO was fully blocked in presence of continuous infusion of a NO* scavenger, carboxy-PTIO (6 mmol/Kg/min), or of the inhibitor of cGMP activation, methylene blue (83 nmol/Kg/min), or of the cyclam-NO precursor, trans-[RuCl(tfins)(cyclam)](tfms) (cyclam-tfms) (500 mmol/Kg/min). The long lasting BP reduction of cyclam-NO can be interpreted in terms of a slower rate of NO* release (k-NO = 2.2 x 10(-3) S(-1) at 35 degrees C) following chemical reduction (E(0') = 0.10 V vs NHE). In summary, cyclam-NO showed an hypotensive effect around 20 times longer than SNP in either normotensive or hypertensive rats, which was completely inhibited by methylene blue or carboxy-PTIO. Continuous infusion of cyclam-tfms completely blocked the hypotensive effect of cyclam-NO by scavenging the NO* released by the reduced cyclam-NO.     

A taxonomic review of Aramides cajaneus (Aves,Gruiformes, Rallidae) with notes on morphological variation in other species of the genus

The taxonomy of the polytypic and wide-ranging Gray-necked Wood-rail, Aramides cajaneus is reviewed, based on external morphology and voice. Throughout its distribution, there is extensive plumage variation, much of it taxonomically uninformative. However, through three informative plumage characters, as well as morphometric and vocal variation, three phylogenetic species were identified within what is today known as Aramides cajaneus, all of which already had available names: Aramides albiventris Lawrence, 1868, from southern Mexico to northeastern Costa Rica, Aramides cajaneus (Statius Müller, 1776) (sensu stricto), from southwestern Costa Rica to Argentina, and Aramides avicenniae Stotz, 1992, from a small section of the coast of southeastern Brazil. Aramides albiventris presents extensive plumage variation, but with no geographic structure. The song of Aramides cajaneus and Aramides avicenniae is strikingly and completely different from the song of Aramides albiventris. A previously unnoticed parapatric pattern of distribution of Aramides cajaneus and its congener Aramides saracura in southeastern Brazil is described, and we clarify that the name Aramides plumbeicollis, included in the synonymy of Aramides albiventris, was first made available in 1892, rather than in 1888 as is widely referred. In addition, plumage variation in Aramides ypecaha, Aramides wolfi, and Aramides mangle is discussed.     

Nanomicellar Formulation of Clotrimazole Improves Its Antitumor Action toward Human Breast Cancer Cells

Background

Although demonstrated as a selective anticancer drug, the clinical use of clotrimazole (CTZ) is limited due to its low solubility in hydrophilic fluids. Thus, we prepared a water-soluble nanomicellar formulation of CTZ (_nCTZ) and tested on the human breast cancer cell line MCF-7 biology.

Methodology/Principal Findings

CTZ was nanoencapsulated in tween 80 micelles, which generated nanomicelles of, approximately, 17 nm of diameter. MCF-7 cells were treated with _nCTZ and unencapsulated DMSO-solubilized drug (_sCTZ) was used for comparison. After treatment, the cells were evaluated in terms of metabolism, proliferation, survival and structure. We found that _nCTZ was more efficient than _sCTZ at inhibiting glycolytic and other cytosolic and mitochondrial enzymes. Moreover, this increased activity was also observed for lactate production, intracellular ATP content, ROS production and antioxidant potential. As a consequence, _nCTZ-treated MCF-7 cells displayed alterations to the plasma membrane, mitochondria and the nucleus. Finally, _nCTZ induced both apoptosis and necrosis in MCF-7 cells.

Conclusions/Significance

MCF-7 cells are more sensible to _nCTZ than to _sCTZ. This was especially evident on regard to antioxidant potential, which is an important cell defense against drugs that affect cell metabolism. Moreover, this water-soluble formulation of CTZ strengths its potential use as an anticancer medicine.     

Fluorescent sensors of siderophores produced by bacterial pathogens

Siderophores are iron-chelating molecules that solubilize Fe³⁺ for microbial utilization and facilitate colonization or infection of eukaryotes by liberating host iron for bacterial uptake. By fluorescently labeling membrane receptors and binding proteins, we created 20 sensors that detect, discriminate, and quantify apo- and ferric siderophores. The sensor proteins originated from TonB-dependent ligand-gated porins (LGPs) of Escherichia coli (Fiu, FepA, Cir, FhuA, IutA, BtuB), Klebsiella pneumoniae (IroN, FepA, FyuA), Acinetobacter baumannii (PiuA, FepA, PirA, BauA), Pseudomonas aeruginosa (FepA, FpvA), and Caulobacter crescentus (HutA) from a periplasmic E. coli binding protein (FepB) and from a human serum binding protein (siderocalin). They detected ferric catecholates (enterobactin, degraded enterobactin, glucosylated enterobactin, dihydroxybenzoate, dihydroxybenzoyl serine, cefidericol, MB-1), ferric hydroxamates (ferrichromes, aerobactin), mixed iron complexes (yersiniabactin, acinetobactin, pyoverdine), and porphyrins (hemin, vitamin B12). The sensors defined the specificities and corresponding affinities of the LGPs and binding proteins and monitored ferric siderophore and porphyrin transport by microbial pathogens. We also quantified, for the first time, broad recognition of diverse ferric complexes by some LGPs, as well as monospecificity for a single metal chelate by others. In addition to their primary ferric siderophore ligands, most LGPs bound the corresponding aposiderophore with ∼100-fold lower affinity. These sensors provide insights into ferric siderophore biosynthesis and uptake pathways in free-living, commensal, and pathogenic Gram-negative bacteria.     

Influence of altitude, latitude and season of collection (Bergmann's rule) on the dimensions of Lutzomyia intermedia (Lutz & Neiva, 1912) (Diptera, Psychodidae, Phlebotominae).

The influence of altitude and latitude on some structure sizes of Lutzomyia intermedia was noted; several structures of insects collected in higher localities were greater, according to Bergmann's rule. This influence was more remarkable in two localities of the State of Espírito Santo, probably due to greater differences in altitude. Comparing insects from different latitudes, more differences were noted in comparisons of insects from low altitude localities than in those of material from higher altitudes. The small number of differences between insects collected in July and in December does not indicate a defined influence of season and temperature on the size of adults. The possible epidemiological implications of these variations are discussed.     

Metabolic Depression and Increased Reactive Oxygen Species Production by Isolated Mitochondria at Moderately Lower Temperatures

Temperature (T) reduction increases lifespan, but the mechanisms are not understood. Because reactive oxygen species (ROS) contribute to aging, we hypothesized that lowering T might decrease mitochondrial ROS production. We measured respiratory response and ROS production in isolated mitochondria at 32, 35, and 37 °C. Lowering T decreased the rates of resting (state 4) and phosphorylating (state 3) respiration phases. Surprisingly, this respiratory slowdown was associated with an increase of ROS production and hydrogen peroxide release and with elevation of the mitochondrial membrane potential, ΔΨ_m. We also found that at lower T mitochondria produced more carbon-centered lipid radicals, a species known to activate uncoupling proteins. These data indicate that reduced mitochondrial ROS production is not one of the mechanisms mediating lifespan extension at lower T. They suggest instead that increased ROS leakage may mediate mitochondrial responses to hypothermia.     

Impact of sewage sludge on the soil bacterial communities by DNA microarray analysis

Sewage sludge has been used as organic manure to replace chemical fertilizer. The aim of this study was to evaluate the effect of doses of sewage sludge on the soil bacterial community by DNA microarray analysis. A microarray phylochip containing 1,560 partial sequences of 16S rRNA from the most common strains of bacteria was developed for bioprospection. Soil plots from an experimental field in Brazil were assessed with or without sludge treatment containing different doses of nitrogen based on that recommended for maize cultivation. The microarray technique was useful for quickly assessing changes in the bacterial communities and a high variation was observed, mainly in soil treated with high doses of sludge. While sludge containing 25 kg N/ha favored an in crease in the number of members in various phyla, on the other hand sludge with the higher dose regarding to 200 kg N/ha caused a reduction in the number of members in almost all phyla. Proteobacteria often dominant in soils was specifically affected. This study highlights the spread of bacteria to new environments and provides direct information about bacterial composition at specific habitats. Our results have shown that bacterial community structure was greatly affected by sludge application.     

Systemic Blockade of Sialylation in Mice with a Global Inhibitor of Sialyltransferases

Sialic acid terminates glycans of glycoproteins and glycolipids that play numerous biological roles in health and disease. Although genetic tools are available for interrogating the effects of decreased or abolished sialoside expression in mice, pharmacological inhibition of the sialyltransferase family has, to date, not been possible. We have recently shown that a sialic acid analog, 2,4,7,8,9-pentaacetyl-3F_ax-Neu5Ac-CO₂Me (3F-NeuAc), added to the media of cultured cells shuts down sialylation by a mechanism involving its intracellular conversion to CMP-3F-NeuAc, a competitive inhibitor of all sialyltransferases. Here we show that administering 3F-NeuAc to mice dramatically decreases sialylated glycans in cells of all tissues tested, including blood, spleen, liver, brain, lung, heart, kidney, and testes. A single dose results in greatly decreased sialoside expression for over 7 weeks in some tissues. Although blockade of sialylation with 3F-NeuAc does not affect viability of cultured cells, its use in vivo has a deleterious “on target” effect on liver and kidney function. After administration of 3F-NeuAc, liver enzymes in the blood are dramatically altered, and mice develop proteinuria concomitant with dramatic loss of sialic acid in the glomeruli within 4 days, leading to irreversible kidney dysfunction and failure to thrive. These results confirm a critical role for sialosides in liver and kidney function and document the feasibility of pharmacological inhibition of sialyltransferases for in vivo modulation of sialoside expression.