Ouabain-induced hypertension is accompanied by increases in endothelial vasodilator factors

The involvement of nitric oxide (NO), prostaglandins, and calcium-dependent potassium channel (K(Ca)) activators on the negative modulation of phenylephrine-induced contractions was evaluated on the isolated aorta and caudal (CAU) artery obtained from rats treated with ouabain for 5 wk to induce hypertension. In ouabain-treated rats, the reactivity to phenylephrine was reduced in the endothelium-intact aorta but not the CAU segments. Endothelial modulation of phenylephrine contraction, as demonstrated by endothelium removal, NO synthase (NOS) inhibition with N(omega)-nitro-L-arginine methyl ester and aminoguanidine, as well as K(Ca) inhibition with tetraethylammonium, was more pronounced in segments from ouabain-treated animals, and here greater effects were seen in the aorta than in CAU. An increased expression of endothelial NOS and neuronal NOS was seen in the aorta after ouabain treatment. In CAU, only endothelial NOS was detected and ouabain treatment did not alter its expression. These results suggest that ouabain-induced hypertension is accompanied by increased NO release derived from endothelial NOS and neuronal NOS and increased release of an endothelial hyperpolarizing factor that presumably opens K(Ca), all of which contribute to the increased negative modulation of the phenylephrine contraction.     

The Putative Mevalonate Diphosphate Decarboxylase from Picrophilus torridus Is in Reality a Mevalonate-3-Kinase with High Potential for Bioproduction of Isobutene

Mevalonate diphosphate decarboxylase (MVD) is an ATP-dependent enzyme that catalyzes the phosphorylation/decarboxylation of (R)-mevalonate-5-diphosphate to isopentenyl pyrophosphate in the mevalonate (MVA) pathway. MVD is a key enzyme in engineered metabolic pathways for bioproduction of isobutene, since it catalyzes the conversion of 3-hydroxyisovalerate (3-HIV) to isobutene, an important platform chemical. The putative homologue from Picrophilus torridus has been identified as a highly efficient variant in a number of patents, but its detailed characterization has not been reported. In this study, we have successfully purified and characterized the putative MVD from P. torridus. We discovered that it is not a decarboxylase per se but an ATP-dependent enzyme, mevalonate-3-kinase (M3K), which catalyzes the phosphorylation of MVA to mevalonate-3-phosphate. The enzyme''s potential in isobutene formation is due to the conversion of 3-HIV to an unstable 3-phosphate intermediate that undergoes consequent spontaneous decarboxylation to form isobutene. Isobutene production rates were as high as 507 pmol min⁻¹ g cells⁻¹ using Escherichia coli cells expressing the enzyme and 2,880 pmol min⁻¹ mg protein⁻¹ with the purified histidine-tagged enzyme, significantly higher than reported previously. M3K is a key enzyme of the novel MVA pathway discovered very recently in Thermoplasma acidophilum. We suggest that P. torridus metabolizes MVA by the same pathway.     

Methyl jasmonate affects phenolic metabolism and gene expression in blueberry (Vaccinium corymbosum)

Blueberry (Vaccinium corymbosum) is a fruit very much appreciated by consumers for its antioxidant potential and health‐promoting traits. Its beneficial potential properties are mainly due to a high content of anthocyanins and their amount can change after elicitation with methyl jasmonate. The aim of this work is to evaluate the changes in expression of several genes, accumulation of phenolic compounds and alterations in antioxidant potential in two different blueberry cultivars (‘Duke’ and ‘Blueray’) in response to methyl jasmonate (0.1 mM). Results showed that 9 h after treatment, the expression of phenylalanine ammonium lyase, chalcone synthase and anthocyanidin synthase genes was stimulated more in the ‘Blueray’ variety. Among the phenols measured an increase was recorded also for epicatechin and anthocyanin concentrations. ‘Duke’ is a richer sourche of anthocyanins compared to ‘Blueray’, treatment with methyl jasmonate promoted in ‘Blueray’ an increase in pigments as well as in the antioxidant potential, especially in fully ripe berries, but treated ‘Duke’ berries had greater levels, which were not induced by methyl jasmonate treatment. In conclusion, methyl jasmonate was, in some cases, an effective elicitor of phenolic metabolism and gene expression in blueberry, though with different intensity between cultivars.     

Predicting haplotype carriers from SNP genotypes in Bos taurus through linear discriminant analysis

Background

SNP (single nucleotide polymorphisms) genotype data are increasingly available in cattle populations and, among other things, can be used to predict carriers of specific haplotypes. It is therefore convenient to have a practical statistical method for the accurate classification of individuals into carriers and non-carriers. In this paper, we present a procedure combining variable selection (i.e. the selection of predictive SNPs) and linear discriminant analysis for the identification of carriers of a haplotype on BTA19 (Bos taurus autosome 19) known to be associated with reduced cow fertility. A population of 3645 Brown Swiss cows and bulls genotyped with the 54K SNP-chip was available for the analysis.

Results

The overall error rate for the prediction of haplotype carriers was on average very low (∼≤1%). The error rate was found to depend on the number of SNPs in the model and their density around the region of the haplotype on BTA19. The minimum set of SNPs to still achieve accurate predictions was 5, with a total test error rate of 1.59.

Conclusions

The paper describes a procedure to accurately identify haplotype carriers from SNP genotypes in cattle populations. Very few misclassifications were observed, which indicates that this is a very reliable approach for potential applications in cattle breeding.     

The influence of the polar head and the hydrophobic chain on the skin penetration enhancement effect of poly(ethylene glycol) derivatives

The effect of a homologue series of nonionic surfactants, namely poly(ethylene glycol) (PEG) fatty acid esters, differing in oxyethylene (PEG 8, PEG 12, and PEG 40) and fatty acid (stearate, mono and di-laurate, and mono and di-oleate) chain lengths, on in vitro skin permeability of ketoprofen (KTP) vehicled in plasters was investigated. The drug diffusion through hairless mouse skin as well as the effect of the surfactant type and strength was studied by Franz diffusion cells and ATR-FTIR spectroscopy. The use of PEG stearate series revealed that the surfactant with the largest polar head, namely PEG 40, was ineffective in enhancing the skin permeation of KTP, independently of the plaster concentrations. The effect of the hydrophobic chain was investigated only by using the shortest oxyethylene chains. The experimental results revealed that the oxyethylene chain length of surfactants appeared to be more influent than the alkyl chain. The prediction of the absorption enhancing capability of these PEG derivatives appeared related to the vehicle other than the proper combination of the number of ethylene oxide groups and alkyl groups.     

Use of dolphins and caimans as bait for Calophysus macropterus (Lichtenstein, 1819) (Siluriforme: Pimelodidae) in the Amazon

A new fishery has been developing in the Amazon that uses dolphin and caiman species as bait to catch piracatinga (Calophysus macropterus), having thus the potential to cause adverse food‐web impacts; however a lack of basic understanding of this fishery is a limitation to the necessary management action. Interviews with fishers and analyses of fishing records in Brazil were used for the study, including harvest methods, types of baits used, commercialization chains, and the rate of increase of piracatinga yields in recent years. Piracatinga fishers are subsistence fishers who harvest piracatinga as a means to alleviate economic constraints when the catch of other species is not profitable or banned due to (reproductive) closed seasons. Harvesting is done with wooden and nylon crates and cages in which whole or pieces of caimans and dolphins are placed to attract the piracatinga, entrapping them. The piracatinga are then sold to intermediate sellers for resale to a few large fish freezing and processing plants for export to Colombia. Annual piracatinga yields in the study area increased at an average rate of 446.5% per year, from 865 kg in 2003 to 23 176 kg in 2009. Because dolphins and caimans comprise various endangered species, the Brazilian government has recently implemented a ban on this fishery, which can be enforced at fish freezing and processing plants. However, there is a danger that such enforcement will lead to the development of a geographically dispersed chain of commercialization and export, such as currently exists for other species including caimans, which would be impossible to control.     

Autonomic innervation of salivary glands in the armadillo,anteater, and sloth (Edentata)

The intraglandular distribution of adrenergic and cholinergic nerve fibers was studied histochemically in the parotid, mandibular, and sublingual glands of six species of edentates belonging to the three families that comprise the order; namely, the Dasypodidae (armadillos), the Myrmecophagidae (anteaters), and the Bradipodidae (sloths). The following histochemical techniques were used: (a) acetylcholinesterase reaction for the demonstration of cholinergic fibers; (b) formaldehyde- and glyoxylic acid-induced fluorescence for the demonstration of adrenergic fibers. In addition, norepinephrine (NE) was assayed fluorimetrically in the mandibular and parotid glands of the armadillo. A network of acetylcholinesterase-positive nerve fibers surrounds the intra- and interlobular ducts and endpieces of all glands; it is of low density in the mandibular and sublingual gland of the sloth, of high density in the sublingual gland of the anteater and of moderate density in the remaining glands. A vascular cholinergic innervation occurs in all salivary glands. Although present around the vessels, adrenergic new fibers were virtually absent from the parenchyma of all glands, even after in vitro incubation of glandular tissue with NE, or after administration of NE to armadillos previously treated with a monoamine oxidase (MAO) inhibitor. Consistent with this fact, the amount of NE present in the parotid and mandibular gland of the armadillo was extremely low. These findings may indicate that the salivary secretion in the edentates is regulated by the parasympathetic rather than by the sympathetic nervous system.     

Post-transcriptional inhibition of ornithine decarboxylase induction by zinc in a difluoromethylornithine resistant cell line

Addition of Zn^2+_ to cell medium inhibited the induction of ornithine decarboxylase (ODC) activity in ODC overproducing L1210-DFMO^r cells. A significant effect was observed at a concentration as low as 0.01 mM, however a more marked inhibition was caused by the addition of 0.1 mM Zn²⁺. The inhibition of the induction of ODC activity was accompanied by a proportional decrease in the content of immunoreactive ODC protein, whereas the level of ODC mRNA, detemined by a solution hybridization RNase protection assay, was not affected signigicantly. Instead, some acceleration of ODC turnover was observed. the addition of 0.1 mM Co²⁺ or Mn²⁺, but not of other divalent metal ions, also inhibited ODC induction; differently from Zn²⁺ however, these metals affected cell viability and/or cell growth. Removal of endogenous Zn²⁺ by a chelator also provoked a strong decrease of ODC induction, which was reversed by Zn²⁺. However, addition of Zn²⁺ in excess of the chelator proved to be markedly inhibitory. These results indicate that both a restricted Zn²⁺ availability and an enhanced presence of the metal can inhibit the induction of ODC in L1210-DFMO^r cells.     

Chitosan antitranspirant activity is due to abscisic acid-dependent stomatal closure

Chitosan (CHT) is a natural compound able to activate the plant own defence machinery against pathogen attacks and to reduce both transpiration and stomatal opening when applied as foliar spray. The data here reported show that CHT-induced antitranspirant activity in bean plants is mediated by ABA, whose level raised over threefold in treated leaves, 24 h after foliar spraying. This is thought to induce partial stomatal closure via a H₂O₂-mediated process, as confirmed by scanning electron microscopy (SEM) and histo-cytochemistry, and, in turn, a decrease of stomatal conductance to water vapor (G_w) and transpiration rate (E), assessed by gas exchange measurements. The relatively high internal CO₂ concentration (C_i) values, suggest the occurrence of a slight decrease in carboxylation efficiency after CHT treatment, which however did not prevail over stomatal limitations. The intrinsic water use efficiency (WUE_i) of CHT treated plants was not statistically different from controls and the maximal photochemical efficiency (F_v/F_m) of PSII was not affected. Moreover, CHT determined a stimulation of the xanthophyll cycle towards de-epoxidation state. On the whole, these results, besides confirming the effectiveness of CHT in reducing plant transpiration, prove that the mechanism underlying this activity differs from that showed by the commercial antitranspirant Vapor Gard^® (VP). In fact, the efficacy of the latter is based on the formation of a thin antitranspirant film over the leaf and not on the reduction of stomatal opening. Finally, suggestions for possible use of the two antitranspirants in different environmental conditions are discussed.