Conservation of key members in the course of the evolution of the insulin signaling pathway

Our understanding of the evolution of the insulin signaling pathway (ISP) is still incomplete. One intriguing unanswered question is the explanation of the emergence of the glucostatic role of insulin in mammals. To find out whether this is due to the development of new sets of signaling transduction elements in these organisms, or to the establishment of new interactions between pre-existing proteins, we rebuilt putative orthologous ISPs in 17 eukaryotic organisms. Then, we computed the conservation of orthologous ISPs at different levels, from sequence similarity of orthologous proteins to co-evolution of interacting domains. We found that the emergence of glucostatic role in mammals can neither be explained by the development of new sets of signaling elements, nor by the establishment of new interactions between pre-existing proteins. The comparison of orthologous IRS molecules indicates that only in mammals have they acquired their complete functionality as efficient recruiters of effector sub-pathways.     

Phylogeographic patterns of steppe species in Eastern Central Europe: a review and the implications for conservation

The phylogeography of species associated with European steppes and extrazonal xeric grasslands is poorly understood. This paper summarizes the results of recent studies on the phylogeography and conservation genetics of animals (20 taxa of beetles, butterflies, reptiles and rodents) and flowering plants (18 taxa) of such, "steppic" habitats in Eastern Central Europe. Most species show a similar phylogeographic pattern: relatively high genetic similarity within regional groups of populations and moderate-to-high genetic distinctiveness of populations from currently isolated regions located in the studied area. This distinctiveness of populations suggests a survival here during glacial maxima, including areas north of the Bohemian Massif-Carpathians arc. Steppic species generally do not follow the paradigmatic patterns known for temperate biota (south-north “contraction–expansion”), but to some extent are similar to those of arctic-alpine taxa. There are three main groups of taxa within Eastern Central Europe that differ in their contemporary distribution pattern, which may reflect historical origin and expansion routes. Present diversity patterns of the studied steppic species suggest that they share a unique genetic signature and distinct assemblages exist in each of the now isolated areas rich in steppic habitats. At least some of these areas probably act as present “interglacial refugia” for steppic species. This study strongly supports the need to protect steppic species throughout their entire ranges in the region, as the continuous destruction of steppic habitats in some areas may lead not only to the disappearance of local populations, but also to the extinction of unique evolutionary units.     

Classifying glycerol dehydratase by its functional residues and purifying selection in its evolution

Glycerol dehydratase (GD) catalyses glycerol reductive conversion to 3-hydroxypropanaldehyde (3-HPA), this being the first step required for the microbial conversion of glycerol to 1, 3 -propanodiol. GD has been functionally characterised to date and two main groups have been determined, one of them being vitamin B(12)-dependent and the other B(12)-independent. GD evolutionary history has been described and an exhaustive analysis made for detecting the functional residues responsible for type I divergence. GD phylogenetic tree topology was seen to be statistically robust and the data indicated strong purifying selection operating on the GD proteins within it. Two clades were indentified, one for vitamin B(12)-dependent and the other for B(12)- independent classes. The ancient hot-pot residues responsible for protein divergency for each clade were also identified. The basic evolutionary biology for GD proteins has been described, thereby opening the way forward for developing rational mutagenesis studies.     

The anti-fibrotic effect of liver growth factor is associated with decreased intrahepatic levels of matrix metalloproteinases 2 and 9 and transforming growth factor beta 1 in bile duct-ligated rats

Liver growth factor (LGF), a mitogen for liver cells, behaves as an anti-fibrotic agent even in extrahepatic sites, but its mechanistic basis is unknown. We aimed to determine the intrahepatic expression pattern of key modulators of liver fibrosis in bile duct-ligated rats (BDL) after injection of LGF. BDL rats received either LGF (4.5 microg/ratXdose, two doses/week, at time 0 or 2 or 5w after operation, depending on the group (BDL+LGF groups, n=20) or saline (BDL+S groups, n=20). Groups were compared in terms of fibrosis (histomorphometry), liver function (aminopyrine breath test), matrix metalloproteinases MMP-2 and MMP-9, transforming growth factor beta 1 (TGF-beta1) and liver endoglin content (Western blotting), and serum tissue inhibitor of metalloproteinases 1 (TIMP-1) levels (ELISA). In BDL+LGF rats, the fibrotic index was significantly lower at 5w, p=0.006, and at 8w, p=0.04, than in BDL+S rats. Liver function values in BDL+LGF rats were higher than those obtained in BDL+S rats (80% at 5w and 79% at 8w, versus 38% and 29%, p<0.01, taking healthy controls as 100%). Notably, in BDL+LGF rats the intrahepatic expression levels of both MMPs were lower at 2w (MMP-2, p=0.03; MMP-9, p=0.05) and 5w (MMP-2, p=0.05, MMP-9, p=0.04). In addition, the hepatic TGF-beta1 level in BDL+LGF rats was lower at 2w (36%, p=0.008), 5w (50%) and 8wk (37%), whereas intrahepatic endoglin expression remained constant in all BDL rats studied. LGF ameliorates liver fibrosis and improves liver function in BDL rats. The LGF-induced anti-fibrotic effect is associated with a decreased hepatic level of MMP-2, MMP-9 and TGF-beta1 in fibrotic rats.     

Ecotypic differentiation in Medicago polymorpha L. along an environmental gradient in central Chile. I. Phenology,biomass production and reproductive patterns

Burr Medic (Medicago polymorpha) is awidespread, polymorphic annual legume of Mediterranean origin, and as aself-reseeding, highly effective nitrogen fixer it has important value invarious dryland farming systems. In central Chile, it is naturalized along a1000 km long environmental gradient, from an arid zone(29°–31° S) to a perhumid one(37°–38°24' S). Comparative and experimental studiesof its ecotypic differentiation patterns along this gradient should be usefulinelucidating the evolutionary forces at work along environmental gradients, andalso for efforts to select annual legumes for pastures and fallow fieldimprovement in mediterranean climate areas. In this first paper of atwo-part series, we present results of two common-environmentexperiments conducted at a subhumid site in central Chile to compare vegetativeand reproductive traits in 69 populations of Burr Medic collected in 1988 and1994. Multivariate analyses based on 11 phenological and agronomic characters,including phenology and biomass accumulation, winter vigour and degree ofhard-seededness, showed that three principal components (PCs)explained nearly 70% of total variability, in both collections. For thelarger collection (1988), the diagram of dispersion for the first twoPCs – phenology, winter vigour and above-ground biomass –provided good discrimination between accessions from arid-semiarid andhumid-perhumid mediterranean zones. Onset of flowering was positivelycorrelated with both latitude and longitude, as well as mean annualprecipitation/potential evapotranspiration (PP/ETP) ofcollecting site; days from first flower to pod ripening was negativelycorrelated with PP/ETP. In contrast to reports from Syria and Sardinia, noclear trends in Burr Medic seed or pod-related traits was found along theChilean gradient, apart from the concentration of spiny pod accessions in thesouthern, more mesic end of the gradient. Winter vigour (according to asemi-quantitative analysis of growth) was greater in accessions fromthe warmer, arid and semiarid zones than in those from cooler, subhumid tohumidones. Both winter vigour and harvest index were negatively correlated with daysto first flower. Applied and evolutionary aspects of the results are brieflydiscussed.     

Disentangling the paradox of insect phenology: are temporal trends reflecting the response to warming?

The strength and direction of phenological responses to changes in climate have been shown to vary significantly both among species and among populations of a species, with the overall patterns not fully resolved. Here, we studied the temporal and spatial variability associated with the response of several insect species to recent global warming. We use hierarchical models within a model comparison framework to analyze phenological data gathered over 40 years by the Japan Meteorological Agency on the emergence dates of 14 insect species at sites across Japan. Contrary to what has been predicted with global warming, temporal trends of annual emergence showed a later emergence day for some species and sites over time, even though temperatures are warming. However, when emergence data were analyzed as a function of temperature and precipitation, the overall response pointed out an earlier emergence day with warmer conditions. The apparent contradiction between the response to temperature and trends over time indicates that other factors, such as declining populations, may be affecting the date phenological events are being recorded. Overall, the responses by insects were weaker than those found for plants in previous work over the same time period in these ecosystems, suggesting the potential for ecological mismatches with deleterious effects for both suites of species. And although temperature may be the major driver of species phenology, we should be cautious when analyzing phenological datasets as many other factors may also be contributing to the variability in phenology.     

In vitro pollination of isolated ovules of Cichorium intybus L.

 A method for the in vitro pollination of chicory (Cichorium intybus L.) ovules was developed. The purpose was to avoid self-incompatibility after in vitro self-pollination of ovules isolated from flower buds before anthesis and from open flower heads. Seedlings were obtained at a low percentage (0.76%), and the results were explained in terms of pollen viability, pollen germination on the ovule, embryogenesis studies and ploidy analysis. Received: 9 April 1997 / Revision received: 23 July 1997 / Accepted: 6 September 1999     

Chimeric contribution of human extended pluripotent stem cells to monkey embryos ex vivo

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