Low dose leflunomide activates PI3K/Akt signalling in erythroleukemia cells and reduces apoptosis induced by anticancer agents

Rheumatoid arthritis (RA) is characterized by persistent joint synovial tissue inflammation. Leflunomide is an immunomodulatory agent that has been approved for treatment of active RA. In the past few years, uses other than RA treatment have appeared. Leflunomide has been reported to show antitumor potential through inhibition of cancer cell proliferation. We thus tested the antiproliferative potential of leflunomide on HEL and K562 erythroleukemia cells. The findings summarized in this report demonstrate for the first time that low dose leflunomide prolonged survival and reduced apoptosis induced by several anticancer agents in erythroleukemia cells. We showed that in treated cells, leflunomide reduced the signalling pathways involved in promoting apoptosis by reducing p38 MAPK and JNK basal activity. On the other hand, leflunomide transiently activated the ERK signalling pathway and induced a sustained activation of Akt. We also showed that leflunomide reduced caspase-3 activity and DNA fragmentation induced by anticancer agents. By using an inhibitory strategy, we showed that inhibition of Akt activation but not ERK abolished the protective effect of leflunomide. Thus our findings suggested that leflunomide reduced apoptosis induced by anticancer agents through PI3K/Akt signalling activation.     

Waterbird mortality in hypersaline environments: the Wyoming trona ponds

Each year hundreds of salt-encrusted waterbirds, mainly Eared Grebes (Podiceps nigricollis Brehm) die at evaporation ponds of the trona (soda ash) industry in southwestern Wyoming. Clinical investigations attributed the mortality to sodium toxicity because high levels of sodium were found in brains of grebes salvaged from the ponds. This was puzzling because natural history information shows that this species resides in saline and hypersaline environments for most of the year. In addition, field observations gave no indication that grebes at the ponds were ingesting salt or salt crystals. Further, the salt glands were not hypertrophied and gavage experiments showed that healthy birds were able to ingest trona pond water without ill effect. Carcasses immersed in pond water, however, attained brain sodium levels far exceeding those considered toxic within a few hours, indicating that the high levels considered as causal probably resulted from post-mortem events. We attribute the mortality to complications of salt encrustation, notably impaired thermoregulation. Other purported cases of sodium toxicity involving encrusted birds at industrial ponds or hypersaline situations may have a similar etiology.     

Landscape genetics of an endangered lemur (Propithecus tattersalli) within its entire fragmented range

Habitat fragmentation may strongly reduce individuals’ dispersal among resource patches and hence influence population distribution and persistence. We studied the impact of landscape heterogeneity on the dispersal of the golden‐crowned sifaka (Propithecus tattersalli), an endangered social lemur species living in a restricted and highly fragmented landscape. We combined spatial analysis and population genetics methods to describe population units and identify the environmental factors which best predict the rates and patterns of genetic differentiation within and between populations. We used non‐invasive methods to genotype 230 individuals at 13 microsatellites in all the main forest fragments of its entire distribution area. Our analyses suggest that the Manankolana River and geographical distance are the primary structuring factors, while a national road crossing the region does not seem to impede gene flow. Altogether, our results are in agreement with a limited influence of forest habitat connectivity on gene flow patterns (except for North of the species’ range), suggesting that dispersal is still possible today among most forest patches for this species. Within forest patches, we find that dispersal is mainly among neighbouring social groups, hence confirming previous behavioural observations.     

Synthesis and PreliminaryIn Vitro Evaluation of Antimycobacterial Activity of New Pyrrolo[1,2-a]quinoxaline-carboxylic Acid Hydrazide Derivatives

New pyrrolo[1,2-a]quinoxaline-2- or -4-carboxylic acid hydrazide derivatives were synthesized from nitroaniline or 1,2-phenylenediamine, and evaluated in vitro for their antimycobacterial activity as part of a TAACF TB screening program. Two compounds 7c and 13 showed an interesting activity at 6.25?μg/mL against Mycobacterium tuberculosis H₃₇Rv, with a 94 and 100 percentage inhibition, respectively.     

IL‐17F co‐ ;expression improves cell growth characteristics and enhances recombinant protein production during CHO cell line engineering

The generation of a high productivity cell line is a critical step in the production of a therapeutic protein. Many innovative engineering strategies have been devised in order to maximize the expression rate of production cells for increased process efficiency. Less effort has focused on improvements to the cell line generation process, which is typically long and laborious when using mammalian cells. Based on unexpected findings when generating stable CHO cell lines expressing human IL‐17F, we studied the benefit of expressing this protein during the establishment of production cell lines. We demonstrate that IL‐17F expression enhances the rate of selection and overall number of selected cell lines as well as their transgene expression levels. We also show that this benefit is observed with different parental CHO cell lines and selection systems. Furthermore, IL‐17F expression improves the efficiency of cell line subcloning processes. IL‐17F can therefore be exploited in a standard manufacturing process to obtain higher productivity clones in a reduced time frame. Biotechnol. Bioeng. 2013; 110: 1153–1163. © 2012 Wiley Periodicals, Inc.     

Arabidopsis thaliana ASN2 encoding asparagine synthetase is involved in the control of nitrogen assimilation and export during vegetative growth

We investigated the function of ASN2, one of the three genes encoding asparagine synthetase (EC 6.3.5.4), which is the most highly expressed in vegetative leaves of Arabidopsis thaliana. Expression of ASN2 and parallel higher asparagine content in darkness suggest that leaf metabolism involves ASN2 for asparagine synthesis. In asn2‐1 knockout and asn2‐2 knockdown lines, ASN2 disruption caused a defective growth phenotype and ammonium accumulation. The asn2 mutant leaves displayed a depleted asparagine and an accumulation of alanine, GABA, pyruvate and fumarate, indicating an alanine formation from pyruvate through the GABA shunt to consume excess ammonium in the absence of asparagine synthesis. By contrast, asparagine did not contribute to photorespiratory nitrogen recycle as photosynthetic net CO₂ assimilation was not significantly different between lines under both 21 and 2% O₂. ASN2 was found in phloem companion cells by in situ hybridization and immunolocalization. Moreover, lack of asparagine in asn2 phloem sap and lowered ¹⁵N flux to sinks, accompanied by the delayed yellowing (senescence) of asn2 leaves, in the absence of asparagine support a specific role of asparagine in phloem loading and nitrogen reallocation. We conclude that ASN2 is essential for nitrogen assimilation, distribution and remobilization (via the phloem) within the plant.     

Cultivation legacies alter soil nutrients and differentially affect plant species performance nearly a century after abandonment

Cultivation legacies affect native vegetation in old fields of the Great Basin, USA for nearly a century after these fields are abandoned. We hypothesized that cultivation lowered soil nutrients and that this legacy would differentially impact plant performance of four representative Great Basin species. To test these hypotheses, we compared soil nutrients (C, N, P, K, Mg and Ca) between two formerly cultivated and adjacent noncultivated sites in two soil series. We then compared the plant growth and foliar nutrient content of an exotic grass (Bromus tectorum L.), two native grasses (Elymus elymoides [Raf.] Swezey and Achnatherum hymenoides [Roem. and Schult.] Barkworth), and a native forb (Sphaeralcea grossulariifolia [Hook. and Arn.] Rydb) grown in these soils in the greenhouse and in the field. Only one sampling site had reduced soil nutrients associated with cultivation legacies, where most of the negative effects on plant performance were found. E. elymoides appeared to be less affected by cultivation legacies than did A. hymenoides, which had a reduced survivorship and 20 % less above-ground biomass in cultivated soils. No species, including B. tectorum, were favored by cultivation. Our findings suggest that cultivation legacies can affect plant performance of different species in different ways and that altered soil nutrients may interact with other abiotic and biotic cultivation legacies in complex ways.