Effects of unfractionated and fractionated heparins on myeloperoxidase activity and interactions with endothelial cells: possible effects on the pathophysiology of equine laminitis

As heparins are sometimes used to prevent equine laminitis, the interactions between equine neutrophil myeloperoxidase (MPO), unfractionated (UFH) and fractionated low molecular weight (LMWH) heparins and digital endothelium have been investigated. The effects of the heparins on purified equine MPO activity were tested by immunocapture followed by enzymatic detection. Endothelium-MPO interactions were assessed by measuring total and active MPO uptake by arterial and venous digital endothelial cells in culture with or without the addition of heparins. A dose-dependent MPO inhibition by UFH and LMWH was seen, with the greatest reduction in MPO activity noted with the highest concentration of LMWH. The MPO capture was greater in arterial cells, but heparins better inhibited MPO capture in venous cells. The activity of cell-bound MPO was almost completely suppressed by the heparins, and no differences were observed between UFH and LMWH. The results confirm the anti-inflammatory properties of heparins and allow a better understanding of the potential role of MPO in laminitis.     

Shedding light on the migratory patterns of the Amazonian goliath catfish,Brachyplatystoma platynemum,using otolith 87Sr/86Sr analyses

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A Marine λ-Oligocarrageenan Inhibits Migratory and Invasive Ability of MDA-MB-231 Human Breast Cancer Cells through Actions on Heparanase Metabolism and MMP-14/MMP-2 Axis

Sugar-based molecules such as heparins or natural heparan sulfate polysaccharides have been developed and widely studied for controlling heparanase (HPSE) enzymatic activity, a key player in extracellular matrix remodelling during cancer pathogenesis. However, non-enzymatic functions of HPSE have also been described in tumour mechanisms. Given their versatile properties, we hypothesized that sugar-based inhibitors may interfere with enzymatic but also non-enzymatic HPSE activities. In this work, we assessed the effects of an original marine λ-carrageenan derived oligosaccharide (λ-CO) we previously described, along with those of its native counterpart and heparins, on cell viability, proliferation, migration, and invasion of MDA-MB-231 breast cancer cells but also of sh-MDA-MB-231 cells, in which the expression of HPSE was selectively downregulated. We observed no cytotoxic and no anti-proliferative effects of our compounds but surprisingly λ-CO was the most efficient to reduce cell migration and invasion compared with heparins, and in a HPSE-dependent manner. We provided evidence that λ-CO tightly controlled a HPSE/MMP-14/MMP-2 axis, leading to reduced MMP-2 activity. Altogether, this study highlights λ-CO as a potent HPSE “modulator” capable of reducing not only the enzymatic activity of HPSE but also the functions controlled by the HPSE levels.     

Joint action of natural and synthetic photosystem II inhibitors

The inhibitory action on photosystem II of four sorgoleone analogues, isolated from Sorghum bicolor (L) Moench, and two synthetic inhibitors, diuron and bentazone, was tested by measuring oxygen evolution of thylakoid membranes. The inhibition of oxygen evolution for mixtures of inhibitors was compared with the Additive Dose Model (ADM). ADM assumes that, at a defined response level, the effect of a mixture of inhibitors can be unambiguously expressed by the potency of either of the inhibitors applied separately. The slope of the logistic dose-response curves differed between the inhibitors; sorgoleone analogues had the steepest and bentazone the shallowest slope. The difference in slopes makes the interpretation of the isoboles less general and may reflect the differences in the interaction between the natural and the synthetic inhibitors with the binding site. The results suggest that there may be some limitation to ADM, namely that compounds with the same site of action might have different response curves if their mechanism of binding is different. The joint action of inhibitors follows ADM at I₅₀ . Therefore, the inhibitors can replace each other in any mixture ratio, based on the relative potencies of the pure inhibitors, without changing each other's effect on oxygen evolution. The joint action at I₂₀ and I₈₀ sometimes diverged from ADM. © 1999 Society of Chemical Industry     

Association between Myeloperoxidase Concentration in Equine Frozen Semen and Post‐Thawing Parameters

Despite improvement of techniques, semen of 20% of stallions remains unfreezable. Recent studies focused on the impact of reactive oxygen species and oxidant enzymes on semen characteristics. Myeloperoxidase (MPO) is a pro‐oxidant enzyme contained in and released by neutrophils during degranulation or after cell lysis. It is responsible for the formation of hypochlorous acid, a strong oxidant agent, which could damage spermatozoa. The aim of this study was to determine the relation between MPO concentration and characteristics of frozen semen from stallions. Thirty‐five straws from different stallions were analysed. Post‐thawing spermatozoal concentration, and progressive and total motility were determined by Computer‐Assisted Semen Analysis. Freezability was determined according to post‐thawing progressive motility (above or below 15%). Percentage of alive spermatozoa and abnormal forms was determined after Eosin–Nigrosin and Diff‐Quick^® staining, respectively. Post‐thawing MPO concentration was measured by enzyme‐linked immunosorbent assay (ELISA). Our study shows that frozen thawed semen contains large amounts of free MPO. We also observed that post‐thawing MPO ELISA assay can be used as an indicator of equine semen freezability. High MPO concentration samples showed lower total and progressive motility. A higher proportion of abnormal head shape associated with acrosome reaction was observed in our late examinations of the high concentration MPO group. Our results show that MPO adversely affects total and progressive motility of equine semen. A negative correlation between normal motile forms and MPO concentration was also observed. The effect of MPO on dead or abnormal forms remains to be precised.     

Phylogenetic characterisation of Onobrychis species with special focus on the forage crop Onobrychis viciifolia Scop.

Sainfoin (Onobrychis viciifolia Scop.) is a perennial forage legume that possesses beneficial properties in the context of sustainable agriculture. In order to initiate a pre-breeding programme, we have assembled a germplasm collection of O. viciifolia from many geographic regions, as well as a collection of Onobrychis species that might be crossed with O. viciifolia to improve its biological and agronomic properties. The objective of this study was to obtain DNA sequences from a representative sample of these accessions in order to characterise their genetic diversity and to assess the robustness of the Onobrychis taxonomy. Because of potential problems with copurification of tannins and polyphenols from Onobrychis leaf tissue, three methods for DNA extraction were assessed and the most appropriate one identified. DNA sequences were obtained for the nuclear internal transcribed spacer region and the trnH-psbA and trnT-trnL intergenic spacers of the chloroplast genome. Neighbour joining trees were constructed and accessions were assigned to operational taxonomic units. The results indicated that there is substantial genetic diversity among Onobrychis species. Redundancies in species nomenclature were identified, as well as possible overlap between some of the sections into which the Onobrychis genus has been divided. A genetic distinction was apparent between O. viciifolia accessions from western Europe and those from eastern Europe and Asia, reflecting a similar division based on agronomic properties. Recognition of these different though related germplasms will be valuable in the design of breeding programmes for the rational improvement of sainfoin as a forage crop for sustainable agriculture.     

Cytological characterisation of the underutilized forage crop Onobrychis viciifolia Scop. and other members of the Onobrychis genus

Sainfoin (Onobrychis viciifolia Scop.) is a perennial forage legume that possesses beneficial properties in the context of sustainable agriculture. A germplasm collection of O. viciifolia was assembled from many geographic regions, as well as a collection of Onobrychis species that might be crossed with O. viciifolia to improve its biological and agronomic properties. The objective of the present study was to obtain cytological information for a broad range of O. viciifolia varieties and related species, in order to confirm and extend previous findings. The study set was 143 accessions of O. viciifolia as well as 34 accessions of 17 other species from the Onobrychis genus. Three of the O. viciifolia accessions were diploid (2n = 2x = 14) and the remainder tetraploid (2n = 4x = 28). Chromosomes were <5 μm in length. Among the other Onobrychis species, roughly half were found to be diploid and the other half tetraploid, with two species, O. arenaria (Kit.) DC. and O. alba subsp. laconica (Orph. ex Boiss.) Hayek, including a mixture of diploid and tetraploid accessions. The basic number of chromosomes was seven, except for O. aequidentata (Sm.) d’Urv. and O. crista-galli Lam., where it was eight. The 2C value for O. viciifolia was estimated at 2.5 pg. These cytological data will be valuable as part of a pre-breeding programme aimed at the eventual production of improved O. viciifolia varieties for use in sustainable agriculture.     

Has global change induced divergent trends in radial growth of Pinus sylvestris and Pinus halepensis at their bioclimatic limit? The example of the Sainte-Baume forest (south-east France)

? It is agreed that climate (precipitation and temperature) influences the distribution of plant species. Near the margins of a species’ natural range, climate becomes limiting to physiological processes. There, climate change may be expected to have a significant impact on tree growth and the species’ ranges may be altered.

? In order to assess what influence climate change could exert on the distribution of pine species at their margin, radial growth trends in ring-width chronologies over the last century were analysed. In the French Mediterranean area where climate change is characterized by increased temperature, forest plots were selected along an altitudinal transect on the north-facing slope of the Sainte-Baume mountain (Bouches-du-Rhône, France) where the ranges of Pinus sylvestris and Pinus halepensis overlap.

? Two growth patterns were identified. For P. halepensis, radial growth has increased at all altitudes indicating that climate change has improved growth conditions of this species near the margin of its ecological range. For P. sylvestris, radial growth has increased only at low altitudes and even decreased at high altitudes.

? It must be deduced that the growth changes observed cannot be generalised either at the species level or at the geographical level and must be interpreted with great caution.