Lipopolysaccharide induces a fibrotic‐like phenotype in endothelial cells

Endothelial dysfunction is crucial in endotoxaemia‐derived sepsis syndrome pathogenesis. It is well accepted that lipopolysaccharide (LPS) induces endothelial dysfunction through immune system activation. However, LPS can also directly generate actions in endothelial cells (ECs) in the absence of participation by immune cells. Although interactions between LPS and ECs evoke endothelial death, a significant portion of ECs are resistant to LPS challenge. However, the mechanism that confers endothelial resistance to LPS is not known. LPS‐resistant ECs exhibit a fibroblast‐like morphology, suggesting that these ECs enter a fibrotic programme in response to LPS. Thus, our aim was to investigate whether LPS is able to induce endothelial fibrosis in the absence of immune cells and explore the underlying mechanism. Using primary cultures of ECs and culturing intact blood vessels, we demonstrated that LPS is a crucial factor to induce endothelial fibrosis. We demonstrated that LPS was able and sufficient to promote endothelial fibrosis, in the absence of immune cells through an activin receptor–like kinase 5 (ALK5) activity–dependent mechanism. LPS‐challenged ECs showed an up‐regulation of both fibroblast‐specific protein expression and extracellular matrix proteins secretion, as well as a down‐regulation of endothelial markers. These results demonstrate that LPS is a crucial factor in inducing endothelial fibrosis in the absence of immune cells through an ALK5‐dependent mechanism. It is noteworthy that LPS‐induced endothelial fibrosis perpetuates endothelial dysfunction as a maladaptive process rather than a survival mechanism for protection against LPS. These findings are useful in improving current treatment against endotoxaemia‐derived sepsis syndrome and other inflammatory diseases.     

Alnus acuminata in dual symbiosis withFrankia and two different ectomycorrhizal fungi (Alpova austroalnicola andAlpova diplophloeus) growing in soilless growth medium

In this study we investigated the capacity of Andean alder (Alnus acuminate Kunth), inoculated withFrankia and two ectomycorrhizal fungi (Alpova austroalnicola Dominguez andAlpova diplophloeus ([Zeller and Dodge] Trappe and Smith), for nodulation and growth in pots of a soilless medium that contained vermiculite or a mixture of ground basalt rock and vermiculite. The seedlings were inoculated withFrankia suspensions prepared from root nodules ofA. Acuminate, followed by inoculation with spores of either one of the twoAlpova species. The seedlings were grown in a greenhouse for 12 months. The seedlings grown in the vermiculite-based growth medium containing large (1-3 mm) basalt particles andAlpova austroalnicola or medium-sized (0.5-1 mm) basalt particles andA. Diplophloeus had the heaviest shoot and root nodule dry weights and abundant ectomycorrhizal colonization. Ectomycorrhizas formed byA. Acuminate withAlpova austroalnicola is described here for the first time. Growth ofAlnus acuminate inoculated with ectomycorrhizal fungi andFrankia in the soilless primary minerals indicates that Andean alder can alter resource supply by tapping an otherwise unavailable nutrient source.     

Improved bioethanol production in an engineered Kluyveromyces lactis strain shifted from respiratory to fermentative metabolism by deletion of NDI1

In this paper, we report the metabolic engineering of the respiratory yeast Kluyveromyces lactis by construction and characterization of a null mutant (Δklndi1) in the single gene encoding a mitochondrial alternative internal dehydrogenase. Isolated mitochondria of the Δklndi1 mutant show unaffected rate of oxidation of exogenous NADH, but no oxidation of matrix NADH; this confirms that KlNdi1p is the only internal NADH dehydrogenase in K. lactis mitochondria. Permeabilized cells of the Δklndi1 mutant do not show oxidation of matrix NADH, which suggests that shuttle systems to transfer the NADH from mitochondrial matrix to cytosol, for being oxidized by external dehydrogenases, are not functional. The Δklndi1 mutation decreases the chronological life span in absence of nutrients. The expression of KlNDI1 is increased by glutathione reductase depletion. The Δklndi1 mutation shifts the K. lactis metabolism from respiratory to fermentative: the Δklndi1 strain shows reduced respiration rate and increased ethanol production from glucose, while it does not grow in non-fermentable carbon sources such as lactate. The biotechnological benefit of the Δklndi1 mutant for bioethanol production from waste cheese whey lactose was proved.     

Development of the inner ear of the brown trout (Salmo trutta fario): I. Gross morphology and sensory cell proliferation

The gross development of the trout inner ear between embryonic and juvenile stages was studied by light microscopy. The otocyst has already formed in 3–4 mm embryos. The semicircular canals begin to separate from the utriculo-saccular cavity in 6 mm embryos, the anterior canal first, then the posterior and the horizontal canal later. The formation of the saccular cavity begins in 7 mm embryos, whereas that of the lagena occurs in 18 mm fry. The first macular primordia appear before the separation of cavities. The anterior and horizontal crests arise from the primordium of the utricular macula, and the posterior crest, macula lagena, and macula neglecta arise from that of the saccular macula. The macula lagena and macula neglecta appear later. The sensory areas of the labyrinth and the number of receptor cells grow continuously between the embryonic and juvenile stages. © 1993 Wiley-Liss, Inc.     

Recreated practices by Mapuche women that strengthen place identity in new urban spaces of residence in Santiago,Chile

The phenomenon of migration to cities by indigenous Mapuche people of Chile is associated with various consequences, such as the loss of ethnic identity and cultural practices. This study aims to describe how ethnic identity is maintained through the recreation of ancestral cultural practices that Mapuche women promote in their families, generating identification to new spaces of residence. This qualitative research draws on analyses of forty-eight interviews conducted with twelve families from four neighbourhoods in Santiago. The study reveals ways in which key traditional Mapuche practices are translated and recreated through the processes of place-referent continuity and place-congruent continuity in new urban areas of residence which in turn express variant forms of ethnic identity and everyday politics of care that extend beyond folkloric notions of rural indigeneity and more static political ideologies of ethno-national autonomy.     

Pigment Epithelium-Derived Factor Is a Survival Factor for Cerebellar Granule Cells in Culture

Abstract: Pigment epithelium-derived factor (PEDF), purified from human fetal retinal pigment epithelium cell culture medium, was shown to potentiate the differentiation of human Y-79 retinoblastoma cells. To investigate potential neurotrophic effects of PEDF on neurons other than those of retinal derivation, we used cultures of cerebellar granule cells. The number of cerebellar granule cells was significantly larger in the presence of PEDF, as demonstrated by an assay for viable cells that uses 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2 H -tetrazolium, inner salt, conversion, by cell count, and by immunocytochemistry. The effect of PEDF showed a dose-response relationship, with a larger effect in chemically defined medium than in serum-containing medium [ED₅₀ = 30 ng/ml (0.70 n M ) in chemically defined medium and 100 ng/ml (2.3 n M ) in serum-containing medium]. PEDF had no effect on incorporation of bromodeoxyuridine (cell proliferation) or on neurofilament content (neurite outgrowth) measured by an enzyme-linked immunoadsorbent assay. These results demonstrate that PEDF has a neurotrophic survival effect on cerebellar granule cells in culture and suggest the possibility that it may affect other CNS neurons as well.     

The foot-and-mouth disease RNA virus as a model in experimental phylogenetics.

Phylogenetic reconstruction methods are subject to two types of limitations: our knowledge about the true history of organisms and the gross simplification implied in the numerical simulation models of the relationships between them. In such a situation, experimental phylogenetics provides a way to assess the accuracy of the phylogenetic reconstruction methods. Nonetheless, this capacity is only feasible for organisms in which replication and mutation rates are high enough to provide valuable data. On the other hand, experimental phylogenetics also provides insights on the main evolutionary processes acting on viral variability under different population dynamics. Our study with the foot-and-mouth disease virus (FMDV) strongly suggests that the phylogenetic reconstruction methods can infer erroneous phylogenies due to nucleotide convergences between isolates belonging to different experimental lineages. We also point out that the diverse evolutionary mechanisms acting in different experimental dynamics generate alterations and change the frequencies of genetic variants, which can lead to the misinterpretation of the real evolutionary history.