TLC-densitometric analysis of artemisinin for the rapid screening of high-producing plantlets of Artemisia annua L

A simple TLC-densitometric technique has been developed for the rapid and accurate analysis of artemisinin in a large number of Artemisia annua plantlets cultured in vitro. This new analytical method is based on the structural conversion of artemisinin on a silica gel layer by ammonia vapour to form 10-azadesoxyartemisinin, a chromophore-containing compound (lambdamax 320 nm) that can be detected by UV-based TLC densitometry. The TLC system was evaluated quantitatively in terms of product stability, precision, accuracy and calibration. Good linearity was obtained in the range of 0.01-0.12 microg artemisinin. The technique appeared to be accurate and sensitive as compared with the complicated pre-column reaction-HPLC technique. Among 90 samples of A. annua plantlets, the artemisinin content in the leaves appeared to be highly variable, ranging from 0.02 to 0.67% w/w dry weight. These results demonstrate that densitometric TLC can be a cheap and simple technique for the accurate screening of high-artemisinin-producing plants.     

Personality‐related differences in response to habitat in Mediterranean blue tits

Habitat‐specific selection pressures have been widely recognized, but whether selection favours different personality types in different habitats has rarely been evaluated. This study aimed to test whether personality‐related differences in annual reproductive success differed between two populations of blue tits (Cyanistes caeruleus) occupying different Mediterranean habitats (oakwood and pinewood). We measured exploration and parental provisioning behaviours and used a path analysis to ask how the interplay between these two behavioural traits affected reproductive success in each of these two habitats. We found that blue tits breeding in the pinewood were slow‐exploring compared to blue tits breeding in the oakwood, suggesting the occurrence of personality‐related differences in settlement, or behavioural plasticity in response to habitat. Exploration behaviour and feeding rates were positively associated, suggesting that they affect each other or that there is an environmental factor affecting both traits simultaneously. Finally, fast explorers were favoured in the pinewood, while there was no selection acting on exploration behaviour in the oak habitat. These findings emphasize the importance of integrating habitat selection, plasticity and personality in the study of behavioural evolution.     

Household energy and recycling of nutrients and carbon to the soil in integrated crop‐livestock farming systems: a case study in Kumbursa village,Central Highlands of Ethiopia

Soil amendment with organic wastes in the Highlands of Ethiopia has been greatly reduced by widespread use of dung cakes and crop residues as fuels. This study assessed the interaction between household energy and recycling of nutrients and carbon to the soil using household survey, focus group discussions, key informant interviews, direct observations and measurements between 2014 and 2015 in Kumbursa village (Central Highlands of Ethiopia). All surveyed households were entirely dependent on biomass fuel for cooking, with production and consumption rates directly related to wealth status, which significantly varied (P < 0.001) among three farm wealth groups (poor, medium and rich). Crop residues and dung cakes accounted for 80(±3)% by energy content and 85(±4)% by dry mass weight of total biomass fuel consumption. Mean losses were 59(±2) kg ha^?1 yr^?1 nitrogen (109(±8) kg yr^?1 per household), 13.9(±0.3) kg ha^?1 yr^?1 phosphorus (26(±2) kg yr^?1 per household), 79(±2) kg ha^?1 yr^?1 potassium (150(±11) kg yr^?1 per household) and 2100(±40) kg ha^?1 yr^?1 organic carbon (3000(±300) kg yr^?1 per household). Rich farmers lost significantly more carbon and nutrients in fuel than farmers in other wealth groups. However, these losses were spread over a larger area, so losses per land area were significantly higher for medium and poor than for rich farmers. This means that the land of poorer farmers is likely to become degraded more rapidly due to fuel limitations than that of rich farmers, so increasing the poverty gap. The estimated financial loss per household due to not using dung and crop residues as organic fertilizer was 162(±8) USSoil amendment with organic wastes in the Highlands of Ethiopia has been greatly reduced by widespread use of dung cakes and crop residues as fuels. This study assessed the interaction between household energy and recycling of nutrients and carbon to the soil using household survey, focus group discussions, key informant interviews, direct observations and measurements between 2014 and 2015 in Kumbursa village (Central Highlands of Ethiopia). All surveyed households were entirely dependent on biomass fuel for cooking, with production and consumption rates directly related to wealth status, which significantly varied (P < 0.001) among three farm wealth groups (poor, medium and rich). Crop residues and dung cakes accounted for 80(±3)% by energy content and 85(±4)% by dry mass weight of total biomass fuel consumption. Mean losses were 59(±2) kg ha^?1 yr^?1 nitrogen (109(±8) kg yr^?1 per household), 13.9(±0.3) kg ha^?1 yr^?1 phosphorus (26(±2) kg yr^?1 per household), 79(±2) kg ha^?1 yr^?1 potassium (150(±11) kg yr^?1 per household) and 2100(±40) kg ha^?1 yr^?1 organic carbon (3000(±300) kg yr^?1 per household). Rich farmers lost significantly more carbon and nutrients in fuel than farmers in other wealth groups. However, these losses were spread over a larger area, so losses per land area were significantly higher for medium and poor than for rich farmers. This means that the land of poorer farmers is likely to become degraded more rapidly due to fuel limitations than that of rich farmers, so increasing the poverty gap. The estimated financial loss per household due to not using dung and crop residues as organic fertilizer was 162(±8) US$ yr^?1. However, this is less than their value as fuels, which was 490(±20) US$ yr^?1. Therefore, farmers will only be persuaded to use these valuable assets as soil improvers if an alternative, cheaper fuel source can be found.     

Accelerated epithelial cell senescence in IPF and the inhibitory role of SIRT6 in TGF-β-induced senescence of human bronchial epithelial cells

Reepithelialization of remodeled air spaces with bronchial epithelial cells is a prominent pathological finding in idiopathic pulmonary fibrosis (IPF) and is implicated in IPF pathogenesis. Recent studies suggest that epithelial senescence is a risk factor for development of IPF, indicating such reepithelialization may be influenced by the acceleration of cellular senescence. Among the sirtuin (SIRT) family, SIRT6, a class III histone deacetylase, has been demonstrated to antagonize senescence. We evaluated the senescence of bronchiolization in association with SIRT6 expression in IPF lung. Senescence-associated β-galactosidase staining and immunohistochemical detection of p21 were performed to evaluate cellular senescence. As a model for transforming growth factor (TGF)-β-induced senescence of abnormal reepithelialization, we used primary human bronchial epithelial cells (HBEC). The changes of SIRT6, p21, and interleukin (IL)-1β expression levels in HBEC, as well as type I collagen expression levels in fibroblasts, were evaluated. In IPF lung samples, an increase in markers of senescence and SIRT6 expression was found in the bronchial epithelial cells lining cystically remodeled air spaces. We found that TGF-β induced senescence in primary HBEC by increasing p21 expression, and, whereas TGF-β also induced SIRT6, it was not sufficient to inhibit cellular senescence. However, overexpression of SIRT6 efficiently inhibited TGF-β-induced senescence via proteasomal degradation of p21. TGF-β-induced senescent HBEC secreted increased amounts of IL-1β, which was sufficient to induce myofibroblast differentiation in fibroblasts. These findings suggest that accelerated epithelial senescence plays a role in IPF pathogenesis through perpetuating abnormal epithelial-mesenchymal interactions, which can be antagonized by SIRT6.     

Evaluation of the role of the endoplasmic reticulum-Golgi transit in the biogenesis of peroxisomal membrane proteins in wild type and peroxisome biogenesis mutant CHO cells

Peroxisomes are thought to be formed by division of pre-existing peroxisomes after the import of newly synthesized proteins. However, it has been recently suggested that the endoplasmic reticulum (ER) provides an alternative de novo mechanism for peroxisome biogenesis in some cells. To test a possible role of the ER-Golgi transit in peroxisome biogenesis in mammalian cells, we evaluated the biogenesis of three peroxisomal membrane proteins (PMPs): ALDRP (adrenoleukodystrophy related protein), PMP70 and Pex3p in CHO cells. We constructed chimeric genes encoding these PMPs and green fluorescent protein (GFP), and transiently transfected them to wild type and mutant CHO cells, in which normal peroxisomes were replaced by peroxisomal membrane ghosts. The expressed proteins were targeted to peroxisomes and peroxisomal ghosts correctly in the presence or absence of Brefeldin A (BFA), a drug known to block the ER-Golgi transit. Furthermore, low temperature did not disturb the targeting of Pex3p-GFP to peroxisomes. We also constructed two chimeric proteins of PMPs containing an ER retention signal "DEKKMP": GFP-ALDRP-DEKKMP and myc- Pex3p-DEKKMP. These proteins were mostly targeted to peroxisomes. No colocalization with an ER maker was found. These results suggest that the classical ER-Golgi pathway does not play a major role in the biogenesis of mammalian PMPs.