Selenium induces a multi‐targeted cell death process in addition to ROS formation

Selenium compounds inhibit neoplastic growth. Redox active selenium compounds are evolving as promising chemotherapeutic agents through tumour selectivity and multi‐target response, which are of great benefit in preventing development of drug resistance. Generation of reactive oxygen species is implicated in selenium‐mediated cytotoxic effects on cancer cells. Recent findings indicate that activation of diverse intracellular signalling leading to cell death depends on the chemical form of selenium applied and/or cell line investigated. In the present study, we aimed at deciphering different modes of cell death in a single cell line (HeLa) upon treatment with three redox active selenium compounds (selenite, selenodiglutathione and seleno‐DL‐cystine). Both selenite and selenodiglutathione exhibited equipotent toxicity (IC₅₀ 5 μM) in these cells with striking differences in toxicity mechanisms. Morphological and molecular alterations provided evidence of necroptosis‐like cell death in selenite treatment, whereas selenodiglutathione induced apoptosis‐like cell death. We demonstrate that selenodiglutathione efficiently glutathionylated free protein thiols, which might explain the early differences in cytotoxic effects induced by selenite and selenodiglutathione. In contrast, seleno‐DL‐cystine treatment at an IC₅₀ concentration of 100 μM induced morphologically two distinct different types of cell death, one with apoptosis‐like phenotype, while the other was reminiscent of paraptosis‐like cell death, characterized by induction of unfolded protein response, ER‐stress and occurrence of large cytoplasmic vacuoles. Collectively, the current results underline the diverse cytotoxic effects and variable potential of redox active selenium compounds on the survival of HeLa cells and thereby substantiate the potential of chemical species‐specific usage of selenium in the treatment of cancers.     

Role of vascular endothelial growth factor in the communication between human osteoprogenitors and endothelial cells

Proper bone remodeling requires an active process of angiogenesis which in turn supplies the necessary growth factors and stem cells. This tissue cooperation suggests a cross‐talk between osteoblasts and endothelial cells. This work aims to identify the role of paracrine communication through vascular endothelial growth factor (VEGF) in co‐culture between osteoblastic and endothelial cells. Through a well defined direct contact co‐culture model between human osteoprogenitors (HOPs) and human umbilical vein endothelial cells (HUVECs), we observed that HUVECs were able to migrate along HOPs, inducing the formation of specific tubular‐like structures. VEGF₁₆₅ gene expression was detected in the HOPs, was up‐regulated in the co‐cultured HOPs and both Flt‐1 and KDR gene expression increased in co‐cultured HUVECs. However, the cell rearrangement observed in co‐culture was promoted by a combination of soluble chemoattractive factors and not by VEGF₁₆₅ alone. Despite having no observable effect on endothelial cell tubular‐like formation, VEGF appeared to have a crucial role in osteoblastic differentiation since the inhibition of its receptors reduced the co‐culture‐stimulated osteoblastic phenotype. This co‐culture system appears to enhance both primary angiogenesis events and osteoblastic differentiation, thus allowing for the development of new strategies in vascularized bone tissue engineering. J. Cell. Biochem. 106: 390–398, 2009. © 2009 Wiley‐Liss, Inc.     

Ligulate Desmarestia (Desmarestiales,Phaeophyceae) revisited: D. japonica sp. nov. and D. dudresnayi differ from D. ligulata

The phylogeny of ligulate and sulfuric‐acid containing species of Desmarestia, occurring worldwide from polar to temperate regions, was revised using a multigenic and polyphasic approach. Sequence data, gametophyte characteristics, and sporophyte morphology support reducing a total of 16 taxa to four different species. (1) D. herbacea, containing broad‐bladed and highly branched forms, has dioecious gametophytes. The three other species have monoecious gametophytes: (2) D. ligulata which is profusely branched and, except for one subspecies, narrow‐bladed, (3) Japanese ligulate Desmarestia, here described as D. japonica sp. nov., which is morphologically similar to D. ligulata but genetically distant from all other ligulate taxa. This species may have conserved the morphology of original ligulate Desmarestia. (4) D. dudresnayi, including unbranched or little branched broad‐bladed taxa. A figure of the holotype of D. dudresnayi, which was lost for decades, was relocated. The taxonomy is complemented by a comparison of internal transcribed spacer and cytochrome c oxidase subunit I (cox1) as potential barcode loci, with cox1 offering good resolution, reflecting species delimitations within the genus Desmarestia.     

Metabolism of D(-)lactic acid in rats given high intragastral doses

In rats given D[u-14C]-labelled DL-lactate with 3.8-13.4 mmol D-lactate per kg 0.75 by stomach tube, the exhalation of CO2 produced from and the renal excretion of D-lactate and metabolites were measured. Exhalation of D-lactate-C accounted for 45-30% of the dosage given with a decreasing proportion at higher doses. The renal excretion of D-lactate averaged 0.9% and that of metabolites from D-lactate 2.4% of the doses given. The fraction of unrecovered D-lactate accounted for about 54-68% of the doses given and increased with doses. The time course of D-lactate oxidation indicated a maximum rate of about 1.5 mmol C per kg 0.75 in 1 hr which was reached at 1 hr after infusion at the earliest and extended up to 8 hr if high doses were given.     

A COMPARATIVE STUDY OF TEMPERATURE RESPONSES OF CARIBBEAN SEAWEEDS FROM DIFFERENT BIOGEOGRAPHIC GROUPS1

Temperature tolerances were determined for Caribbean isolates (total 31) of seaureds belonging to three distributional groups: 1) species confined to the tropical western Atlantic (Botryocladia spinulifera, Chamaedoris peniculum, Cladophoropsis sundanensis, Dictyopteris justii, Dictyurus occidentalis, Haloplegma duperreyi, and Heterosiphonia gibbesii); 2) amphi-Atlantic species with a (sub)tropical distribution that have their northern boundary in the eastern Atlantic at the tropical Cape Verde Islands (Bryothamnion triquetrum and Ceramium nitens) or the subtropical Canary Islands (Ceratodictyon intricatum, Coelothrix irregularis, Dictyopteris delicatula, Ernodesmis verticillata, and Lophocladia trichoclados; and 3) species with an am-phi-Atlantic tropical to warm-temperate distribution also occurring in the Mediterranean (Cladophoropsis membranacea, Digenea simplex, Microdictyon boergesenii, and Wurdemannia miniata). For some isolates, growth response curves and temperature requirements for reproduction were also determined. Growth occurred in the range (18)20–30° C with optimum growth rates at 25°–30°C, irrespective of distribution group. Reproduction generally occurred at (20)25°–30° C although there were some exceptions. Species were extremely stenothermal, with those restricted to the western Atlantic surviving a total range of only 10/13° C (between 18/20° and 30/33° C). Tolerance to high temperatures was correlated with vertical position in the iniertidal/subtidal zone rather than biogeography grouping. Species restricted to the subtidal were the least tolerant, with permanent survival at 30° C but not at 33°C. Tolerance to low temperatures was not different in subtidal and intertidal species but was significantly better in am phi-Atlantic than in western Atlantic species. In the former group, damage occurred at 15°–18° C but in the latter group at 18°-20° C. We propose that these differences in low-temperature tolerances in Caribbean populations of species from different distribution groups reflect adaptations to glacial cold-stress in the tropical eastern Atlantic and subsequent trans-Atlantic dispersal.     

Genotypic variations in GST genes reveal a regulatory role in the accumulation of caffeoylquinic acids in leafy sweet potato (Ipomoea batatas)

Plant Cell, Tissue and Organ Culture (PCTOC) - Leafy sweet potato is rich in caffeoylquinic acids (CQAs), including monoCQAs and diCQAs, which play important roles in plant resistance to...     

Chemical Composition and Spectrum of Insecticidal Activity of the Essential Oils of Ocimum gratissimum L. and Cymbopogon citratus Stapf on the Main Insects of the Cotton Entomofauna in Côte d'Ivoire

Among the alternatives to environmentally toxic and socio-economically unacceptable chemical pesticides, essential oils from Ocimum gratissimum and Cymbopogon citratus were tested on the main pests and beneficial insects of the cotton plant in Côte d′Ivoire. After extraction and chemical analysis of the essential oils, field trials were carried out using a Fisher block system with three treatment repetitions where their effects compared with those of a registered synthetic insecticide (IBIS A 52 EC). Foliar applications of the products were carried out in accordance with the cotton plant protection extension programme in Côte d′Ivoire from the 45th to the 115th day after plant emergence, with one application every fortnight. Twenty-three and forty compounds representing about 96 and 99 % of the oil composition of O. gratissimum and C. citratus respectively were elucidated. The most abundant compounds were p-cymene and thymol (O. gratissimum) and myrcene, neral and geranial (C. citratus). The essential oil of O. gratissimum at concentrations of 2 and 5 % showed insecticidal activity on all pests (biting-sucking and carpophagous), except the phyllophagous Syllepte derogata. C. citratus, at a low concentration (1 %), was particularly toxic to whiteflies (Bemisia tabaci), however, it favoured the action of beneficial insects, specifically black ants and ladybirds in the cotton plots, unlike the chemical product. EO of O. gratissimum (1.60 and 4.62 mg GALAE/g, respectively) and C. citratus (2.26 and 2.78 mg GALAE, respectively) exhibited also significant acetyl and butyryl cholinesterase inhibitors. Insecticide formulations based on the essential oils of O. gratissimum and C. citratus offer favourable prospects for their use in cotton cultivation as an alternative to chemical pesticides.     

uPA and MMP‐2 were involved in self‐assembled network formation in a two dimensional co‐culture model of bone marrow stromal cells and endothelial cells

Two dimensional (2D) co‐cultures of human bone marrow stromal cells (HBMSCs) and human umbilical vein endothelial cells (HUVECs) stimulate osteoblastic differentiation of HBMSCs, induce the formation of self‐assembled network and cell interactions between the two cell types involving many vascular molecules. Because of their strong activities on angiogenesis and tissue remodeling, urokinase plasminogen activator (uPA), plasminogen activator inhibitor‐1 (PAI‐1), matrix metalloproteinase‐2 (MMP‐2) as well tissue inhibitors of matrix metalloproteinase‐2 (TIMP‐2) were investigated in this 2D co‐culture model. We found that the expression of uPA, MMP‐2 in the co‐cultured cells was significantly higher than those in mono‐cultured cells. In opposite, PAI‐1, expressed only by HUVECs is not regulated in the co‐culture. Inhibition assays confirm that uPA played a critical role in the formation of self‐assembled network as neutralization of uPA disturbed this network. In the same context, inhibition of MMP‐2 prevented the formation of self‐assembled network, while the inhibition of uPA abolished the over expression and the activity of MMP‐2. This upregulation could initiate the uPA expression and proteolysis processes through the MMP‐2 activity, and may contribute to endothelial cell migration and the formation of this self‐assembled network observed in these 2D co‐cultured cells. J. Cell. Biochem. 114: 650–657, 2013. © 2012 Wiley Periodicals, Inc.     

Quaternary Structure of Pathological Prion Protein as a Determining Factor of Strain-Specific Prion Replication Dynamics

Prions are proteinaceous infectious agents responsible for fatal neurodegenerative diseases in animals and humans. They are essentially composed of PrP^Sc, an aggregated, misfolded conformer of the ubiquitously expressed host-encoded prion protein (PrP^C). Stable variations in PrP^Sc conformation are assumed to encode the phenotypically tangible prion strains diversity. However the direct contribution of PrP^Sc quaternary structure to the strain biological information remains mostly unknown. Applying a sedimentation velocity fractionation technique to a panel of ovine prion strains, classified as fast and slow according to their incubation time in ovine PrP transgenic mice, has previously led to the observation that the relationship between prion infectivity and PrP^Sc quaternary structure was not univocal. For the fast strains specifically, infectivity sedimented slowly and segregated from the bulk of proteinase-K resistant PrP^Sc. To carefully separate the respective contributions of size and density to this hydrodynamic behavior, we performed sedimentation at the equilibrium and varied the solubilization conditions. The density profile of prion infectivity and proteinase-K resistant PrP^Sc tended to overlap whatever the strain, fast or slow, leaving only size as the main responsible factor for the specific velocity properties of the fast strain most infectious component. We further show that this velocity-isolable population of discrete assemblies perfectly resists limited proteolysis and that its templating activity, as assessed by protein misfolding cyclic amplification outcompetes by several orders of magnitude that of the bulk of larger size PrP^Sc aggregates. Together, the tight correlation between small size, conversion efficiency and duration of disease establishes PrP^Sc quaternary structure as a determining factor of prion replication dynamics. For certain strains, a subset of PrP assemblies appears to be the best template for prion replication. This has important implications for fundamental studies on prions.