The A to I editing landscape in melanoma and its relation to clinical outcome

RNA editing refers to non-transient RNA modifications that occur after transcription and prior to translation by the ribosomes. RNA editing is more widespread in cancer cells than in non-transformed cells and is associated with tumorigenesis of various cancer tissues. However, RNA editing can also generate neo-antigens that expose tumour cells to host immunosurveillance. Global RNA editing in melanoma and its relevance to clinical outcome currently remain poorly characterized. The present study compared RNA editing as well as gene expression in tumour cell lines from melanoma patients of short or long metastasis-free survival, patients relapsing or not after immuno- and targeted therapy and tumours harbouring BRAF or NRAS mutations. Overall, our results showed that NTRK gene expression can be a marker of resistance to BRAF and MEK inhibition and gives some insights of candidate genes as potential biomarkers. In addition, this study revealed an increase in Adenosine-to-Inosine editing in Alu regions and in non-repetitive regions, including the hyperediting of the MOK and DZIP3 genes in relapsed tumour samples during targeted therapy and of the ZBTB11 gene in NRAS mutated melanoma cells. Therefore, RNA editing could be a promising tool for identifying predictive markers, tumour neoantigens and targetable pathways that could help in preventing relapses during immuno- or targeted therapies

Spatio-temporal distribution of pyrethroids in soil in Mediterranean paddy fields

[EN] The demand of rice by the increase in population in many countries has intensified the application of pesticides and the use of poor quality water to irrigate fields. The terrestrial environment is one compartment affected by these situations, where soil is working as a reservoir, retaining organic pollutants. Therefore, it is necessary to develop methods to determine insecticides in soil and monitor susceptible areas to be contaminated, applying adequate techniques to remediate them. Materials and methods This study investigates the occurrence of ten pyrethroid insecticides (PYs) and its spatio-temporal variance in soil at two different depths collected in two periods (before plow and during rice production), in a paddy field area located in the Mediterranean coast. Pyrethroids were quantified using gas chromatography mass spectrometry (GC MS) after ultrasound-assisted extraction with ethyl acetate. The results obtained were assessed statistically using non-parametric methods, and significant statistical differences (p&#8201;<&#8201;0.05) in pyrethroids content with soil depth and proximity to wastewater treatment plants were evaluated. Moreover, a geographic information system (GIS) was used to monitor the occurrence of PYs in paddy fields and detect risk areas. Results and discussion Pyrethroids were detected at concentrations &#8804;57.0 ng g&#8722;1 before plow and &#8804;62.3 ng g&#8722;1 during rice production, being resmethrin and cyfluthrin the compounds found at higher concentrations in soil. Pyrethroids were detected mainly at the top soil, and a GIS program was used to depict the obtained results, showing that effluents from wastewater treatment plants (WWTPs) were the main sources of soil contamination. No toxic effects were expected to soil organisms, but it is of concern that PYs may affect aquatic organisms, which represents the worst case scenario. Conclusions A methodology to determine pyrethroids in soil was developed to monitor a paddy field area. The use of water from WWTPs to irrigate rice fields is one of the main pollution sources of pyrethroids. It is a matter of concern that PYs may present toxic effects on aquatic organisms, as they can be desorbed from soil. Phytoremediation may play an important role in this area, reducing the possible risk associated to PYs levels in soil.Authors wish to thank INIA for the predoctoral fellowship (R. Aznar) and Spanish Ministry of Economy and Competitiveness RTA2014-00012-C03-01 for financial support and Jonathan Villanueva Martin for his contribution to this work.Aznar, R.; Moreno-Ramón, H.; Albero, B.; Sánchez Brunete, C.; Tadeo, JL. (2016). Spatio-temporal distribution of pyrethroids in soil in Mediterranean paddy fields. Journal of Soils and Sediments. 17(5):1503-1513. https://doi.org/10.1007/s11368-016-1417-2S15031513175Albaseer SS, Rao RN, Swamy YV, Mukkanti K (2010) An overview of sample preparation and extraction of synthetic pyrethroids from water, sediment and soil. J Chromatogr A 1217(35):5537–5554Alonso MB, Feo ML, Corcellas C, Vidal LG, Bertozzi CP, Marigo J, Secchi ER, Bassoi M, Azevedo AF, Dorneles PR, Torres JPM, Lailson-Brito J, Malm O, Eljarrat E, Barcelo D (2012) Pyrethroids: a new threat to marine mammals? Environ Int 47:99–106Amweg EL, Weston DP, Ureda NM (2005) Use and toxicity of pyrethroid pesticides in the Central Valley, California, USA. Environ Toxicol Chem 24(4):966–972Arias-Estevez M, Lopez-Periago E, Martinez-Carballo E, Simal-Gandara J, Mejuto JC, Garcia-Rio L (2008) The mobility and degradation of pesticides in soils and the pollution of groundwater resources. Agric Eco Environ 123(4):247–260Aznar R, Albero B, Sanchez-Brunete C, Miguel E, Tadeo JL (2014) Multiresidue analysis of insecticides and other selected environmental contaminants in poultry manure by gas chromatography/mass spectrometry. J AOAC Int 97(4):978–986Campo J, Masia A, Blasco C, Pico Y (2013) Occurrence and removal efficiency of pesticides in sewage treatment plants of four Mediterranean River Basins. J Hazard Mater 263:146–157European Commission (2002) Review report for the active substance Cyfluthrin, 6843/VI/97-finalEuropean Commission (2004) Review report for the active substance α-Cypermethrin, SANCO/4335/2000-finalEuropean Commission (2005) Review report for the active substance Esfenvalerate, 6846/VI/97-finalFeo ML, Ginebreda A, Eljarrat E, Barcelo D (2010) Presence of pyrethroid pesticides in water and sediments of Ebro River Delta. J Hydrol 393(3-4):156–162Fojut TL, Palumbo AJ, Tjeerdema RS (2012) Aquatic life water quality criteria derived via the UC Davis method: II. Pyrethroid insecticides. Rev Environ Contam Toxicol 216:51–103Gan J, Lee SJ, Liu WP, Haver DL, KAbashima JN (2005) Distribution and persistence of pyrethroids in runoff sediments. J Environ Qual 34:836–841Hill IR (1985) Aquatic organisms and pyrethroids. Pestic Sci 27:429–465Huang LM, Thompson A, Zhang GL, Chen LM, Han GZ, Gong ZT (2015) The use of chronosequences in studies of paddy soil evolution: a review. Geoderma 237:199–210Katagi T (2004) Photodegradation of pesticides on plant and soil surfaces. Rev Environ Contam Toxicol 182:1–189Laskowski DA (2002) Physical and chemical properties of pyrethroids. Rev Environ Contam Toxicol 174:49–170Mahabali S, Spagnoghe P (2014) Mitigation of two insecticides by wetlands plants: feasibility study for the treatment of agricultural runoff in Suriname (South America). Water Air Soil Pollut 225:1771Maund SJ, Hamer MJ, Lane MCG, Farrelly E, Rapley JH, Goggin UM, Gentle WE (2002) Partitioning, bioavailability, and toxicity of the pyrethroid insecticide cypermethrin in sediments. Environ Toxicol Chem 21(1):9–15Maund SJ, Campbell PJ, Giddings JM, Hamer MJ, Henry K, Pilling ED, Warinton JS, Wheeler JR (2012) Ecotoxicology of synthetic pyrethroids. Top Curr Chem 314:137–165Money E, Carter GP, Serre ML (2009) Using river distances in the space/time estimation of dissolved oxygen along two impaired river networks in New Jersey. Water Res 43(7):1948–1958Moore MT, Cooper CM, Smith S, Jr Cullum RF, Knight SS, Locke MA, Bennett ER (2009) Mitigation of two pyrethroid insecticides in Mississippi Delta constructed wetland. Environ Pollut 157:250–256Moreno-Ramón H, Marqués-Mateu A, Ibáñez-Asensio S, Gisbert JM (2015) Wetland soils under rice management and seawater intrusion: characterization and classification. Spa J Soil Sci 5(2):111–129Nawaz MF, Bourrie G, Trolard F, Mouret JC, Henry P (2013) Effects of agronomic practices on the physico-chemical properties of soil waters in rice culture. Turk J Agric For 37(2):195–202Oros DR, Werner I (2005) Pyrethroid insecticides: an analysis of use patterns, distributions, potential toxicity and fate in the Sacramento-San Joaquin Delta and Central Valley. White Paper for the Interagency Ecological Program. SFEI Contribution 415. San Francisco Estuary Institute, Oakland, CAPascual-Aguilar J, Andreu V, Gimeno-Garcia E, Pico Y (2015) Current anthropogenic pressures on agro-ecological protected coastal wetlands. Sci Total Environ 03:190–199Soil Survey Staff (2014a) Soil survey field and laboratory methods manual. Soil survey investigations report no. 51, version 2.0. In: Burt R, Soil Survey Staff (eds). U.S. Department of Agriculture, Natural Resources Conservation Service, Washington, p 407Soil Survey Staff (ed) (2014b) Keys to soil taxonomy, 12th edn. USDA-Natural Resources Conservation Service, Washington, p 372Song Y, Kai J, Song X, Zhang W, Li L (2015) Long-term toxic effects of deltamethrin and fenvalerate in soil. J Hazard Mater 289:158–164Weston DP, Holmes RW, You J, Lydy MJ (2005) Aquatic toxicity due to residential use of pyrethroid insecticides. Environ Sci Technol 39(24):9778–9784Weston DP, Ramil HL, Lydy MJ (2013) Pyrethroid insecticides in municipal wastewater. Environ Toxicol Chem 32(11):2460–2468Zhou JL, Rowland S, Mantoura RFC (1995) Partition of synthetic pyrethroid insecticides between dissolved and particulate phases. Water Res 29:1023–110

Agouti Revisited: Transcript Quantification of the ASIP Gene in Bovine Tissues Related to Protein Expression and Localization

Beside its role in melanogenesis, the agouti signaling protein (ASIP) has been related to obesity. The potentially crucial role in adipocyte development makes it a tempting candidate for economic relevant, fat related traits in farm animals. The objective of our study was to characterize the mRNA expression of different ASIP transcripts and of putative targets in different bovine tissues, as well as to study consequences on protein abundance and localization. ASIP mRNA abundance was determined by RT-qPCR in adipose and further tissues of cattle representing different breeds and crosses. ASIP mRNA was up-regulated more than 9-fold in intramuscular fat of Japanese Black cattle compared to Holstein (p<0.001). Further analyses revealed that a transposon-derived transcript was solely responsible for the increased ASIP mRNA abundance. This transcript was observed in single individuals of different breeds indicating a wide spread occurrence of this insertion at the ASIP locus in cattle. The protein was detected in different adipose tissues, skin, lung and liver, but not in skeletal muscle by Western blot with a bovine-specific ASIP antibody. However, the protein abundance was not related to the observed ASIP mRNA over-expression. Immuno-histochemical analyses revealed a putative nuclear localization of ASIP additionally to the expected cytosolic signal in different cell types. The expression of melanocortin receptors (MCR) 1 to 5 as potential targets for ASIP was analyzed by RT-PCR in subcutaneous fat. Only MC1R and MC4R were detected indicating a similar receptor expression like in human adipose tissue. Our results provide evidence for a widespread expression of ASIP in bovine tissues at mRNA and, for the first time, at protein level. ASIP protein is detectable in adipocytes as well as in further cells of adipose tissue. We generated a basis for a more detailed investigation of ASIP function in peripheral tissues of various mammalian species

Expression of melanocortin receptors mRNA, and direct effects of ACTH on steroids secretion in the bovine ovary

Melanocortin receptors (MCRs) are involved in physiological responses to ACTH, as well as to α-, β- and γ-melanocyte-stimulating hormone (α-, β- and γ-MSH). Their expression has previously been analyzed in various bovine tissues; however, there are apparently no reports regarding their localization in the ovaries. In the present study, the expression of MCR mRNA in various bovine ovarian structures was characterized with reverse transcription polymerase chain reaction (RT-PCR). Furthermore, whether ACTH affected follicular components by affecting steroid secretion in fragments of ovarian follicular wall of medium and large antral follicles cultured in serum free medium with 1, 10, and 100 nM ACTH, was also determined. Melanocortin receptors mRNA was localized in the theca cells of various follicular stages, whereas only MC3R mRNA was weakly evident in granulosa cells. Melanocortin receptors 1, 2, and 3 mRNA were present in the CL, whereas in stroma, only MC2R mRNA was expressed. There were significant increases in estradiol and cortisol concentrations in response to ACTH in medium follicles, as well as increased concentrations of testosterone and cortisol in large follicles. These results confirmed earlier reports in other species, and demonstrated that MCRs were present in bovine ovaries. Since ACTH induced steroid secretion from the ovary in vitro, we inferred that melanocortin peptides could be involved in regulatory mechanisms related to ovarian functions, e.g. ovulation, steroidogenesis, and luteal function.Fil: Amweg, Ayelen Noelia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; ArgentinaFil: Alfonso, Paredes. Universidad de Chile; ChileFil: Salvetti, Natalia Raquel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; ArgentinaFil: Lara, Hernan E.. Universidad de Chile; ChileFil: Ortega, Hugo Hector. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentin

The A to I editing landscape in melanoma and its relation to clinical outcome.

RNA editing refers to non-transient RNA modifications that occur after transcription and prior to translation by the ribosomes. RNA editing is more widespread in cancer cells than in non-transformed cells and is associated with tumorigenesis of various cancer tissues. However, RNA editing can also generate neo-antigens that expose tumour cells to host immunosurveillance. Global RNA editing in melanoma and its relevance to clinical outcome currently remain poorly characterized. The present study compared RNA editing as well as gene expression in tumour cell lines from melanoma patients of short or long metastasis-free survival, patients relapsing or not after immuno- and targeted therapy and tumours harbouring BRAF or NRAS mutations. Overall, our results showed that NTRK gene expression can be a marker of resistance to BRAF and MEK inhibition and gives some insights of candidate genes as potential biomarkers. In addition, this study revealed an increase in Adenosine-to-Inosine editing in Alu regions and in non-repetitive regions, including the hyperediting of the MOK and DZIP3 genes in relapsed tumour samples during targeted therapy and of the ZBTB11 gene in NRAS mutated melanoma cells. Therefore, RNA editing could be a promising tool for identifying predictive markers, tumour neoantigens and targetable pathways that could help in preventing relapses during immuno- or targeted therapies