COT drives resistance to RAF inhibition through MAP kinase pathway reactivation

Oncogenic mutations in the serine/threonine kinase B-RAF (also known as BRAF) are found in 50-70% of malignant melanomas. Pre-clinical studies have demonstrated that the B-RAF(V600E) mutation predicts a dependency on the mitogen-activated protein kinase (MAPK) signalling cascade in melanoma-an observation that has been validated by the success of RAF and MEK inhibitors in clinical trials. However, clinical responses to targeted anticancer therapeutics are frequently confounded by de novo or acquired resistance. Identification of resistance mechanisms in a manner that elucidates alternative 'druggable' targets may inform effective long-term treatment strategies. Here we expressed ～600 kinase and kinase-related open reading frames (ORFs) in parallel to interrogate resistance to a selective RAF kinase inhibitor. We identified MAP3K8 (the gene encoding COT/Tpl2) as a MAPK pathway agonist that drives resistance to RAF inhibition in B-RAF(V600E) cell lines. COT activates ERK primarily through MEK-dependent mechanisms that do not require RAF signalling. Moreover, COT expression is associated with de novo resistance in B-RAF(V600E) cultured cell lines and acquired resistance in melanoma cells and tissue obtained from relapsing patients following treatment with MEK or RAF inhibitors. We further identify combinatorial MAPK pathway inhibition or targeting of COT kinase activity as possible therapeutic strategies for reducing MAPK pathway activation in this setting. Together, these results provide new insights into resistance mechanisms involving the MAPK pathway and articulate an integrative approach through which high-throughput functional screens may inform the development of novel therapeutic strategies.     

The amount of carbon released from peat and forest fires in Indonesia during 1997

Tropical peatlands are one of the largest near-surface reserves of terrestrial organic carbon, and hence their stability has important implications for climate change. In their natural state, lowland tropical peatlands support a luxuriant growth of peat swamp forest overlying peat deposits up to 20 metres thick. Persistent environmental change-in particular, drainage and forest clearing-threatens their stability, and makes them susceptible to fire. This was demonstrated by the occurrence of widespread fires throughout the forested peatlands of Indonesia during the 1997 El Ni?o event. Here, using satellite images of a 2.5 million hectare study area in Central Kalimantan, Borneo, from before and after the 1997 fires, we calculate that 32% (0.79 Mha) of the area had burned, of which peatland accounted for 91.5% (0.73 Mha). Using ground measurements of the burn depth of peat, we estimate that 0.19-0.23 gigatonnes (Gt) of carbon were released to the atmosphere through peat combustion, with a further 0.05 Gt released from burning of the overlying vegetation. Extrapolating these estimates to Indonesia as a whole, we estimate that between 0.81 and 2.57 Gt of carbon were released to the atmosphere in 1997 as a result of burning peat and vegetation in Indonesia. This is equivalent to 13-40% of the mean annual global carbon emissions from fossil fuels, and contributed greatly to the largest annual increase in atmospheric CO(2) concentration detected since records began in 1957 (ref. 1).     

A simple procedure for the photometric evaluation of incidental findings on the recording strips of the Burkard pollen-and-spore trap

Summary Reference is made to a procedure for evaluating certain incidental findings presented by particles of soot or dust on the adhesive strips of the Burkard pollen-and-spore trap or the slides of the similar Hirst apparatus. For this an evaluating device is recommended that is generally used for pherograms in large hospitals today. The often very noticeable streaks of soot or dust particles on the adhesive layer can be assessed with this method. Furthermore, it is possible to determine the time of the occurrence of the particles and possibly to relate them to the velocity and direction of the wind, and also eventually to detect the source of the polluting material.Acknowledgments. We would like to thank Prof. R. Schuppli, director of the Dermatological Department of the Basel Cantonal Hospital, for making it possible to carry out extended monitoring of airborne particles. Our special thanks go to Dr H.P. Rieder, head of the protein-testing laboratory of the Basel Cantonal Hospital, for enabling the examinations with the pherogram evaluator to be made. Finally, we wish to thank the Swiss National Foundation for the Advancement of Scientific Research who has made a grant-in-aid available to Dr R.M. Leuschner for monitoring airborne particles.     

Does prolactin control the blood progesterone level on early dioestrus in rats?

Summary Bromocriptine treatment on either prooestrus or oestrus in female rats did not affect luteal function on the day of dioestrus 1.This investigation was partially financed by the C.N.R.S. (E.R.A. No. 566).Acknowledgments. We are thankful to Mrs C. Lazarus for her excellent technical assistance, to Mr R. Dujol for the figures. We wish to express our gratitude to Dr A.F. Parlow for providing reagents for prolactin RIA and to Sandoz Laboratories for bromocriptine. Reprint request should be addressed to Cl.A.     

A thymus candidate in lampreys

Immunologists and evolutionary biologists have been debating the nature of the immune system of jawless vertebrates--lampreys and hagfish--since the nineteenth century. In the past 50 years, these fish were shown to have antibody-like responses and the capacity to reject allografts but were found to lack the immunoglobulin-based adaptive immune system of jawed vertebrates. Recent work has shown that lampreys have lymphocytes that instead express somatically diversified antigen receptors that contain leucine-rich-repeats, termed variable lymphocyte receptors (VLRs), and that the type of VLR expressed is specific to the lymphocyte lineage: T-like lymphocytes express type A VLR (VLRA) genes, and B-like lymphocytes express VLRB genes. These clonally diverse anticipatory antigen receptors are assembled from incomplete genomic fragments by gene conversion, which is thought to be initiated by either of two genes encoding cytosine deaminase, cytosine deaminase 1 (CDA1) in T-like cells and CDA2 in B-like cells. It is unknown whether jawless fish, like jawed vertebrates, have dedicated primary lymphoid organs, such as the thymus, where the development and selection of lymphocytes takes place. Here we identify discrete thymus-like lympho-epithelial structures, termed thymoids, in the tips of the gill filaments and the neighbouring secondary lamellae (both within the gill basket) of lamprey larvae. Only in the thymoids was expression of the orthologue of the gene encoding forkhead box N1 (FOXN1), a marker of the thymopoietic microenvironment in jawed vertebrates, accompanied by expression of CDA1 and VLRA. This expression pattern was unaffected by immunization of lampreys or by stimulation with a T-cell mitogen. Non-functional VLRA gene assemblies were found frequently in the thymoids but not elsewhere, further implicating the thymoid as the site of development of T-like cells in lampreys. These findings suggest that the similarities underlying the dual nature of the adaptive immune systems in the two sister groups of vertebrates extend to primary lymphoid organs.