Highly variable AFLP and S-SAP markers for the identification of ‘Malbec’ and ‘Syrah’ clones

The retrotransposon-based sequence-specific amplification polymorphism (S-SAP) and the amplified fragment length polymorphism (AFLP) marker systems were used to assess the clonal variation of 14 ‘Syrah’ and 22 ‘Malbec’ (Vitis vinifera L.) clones. The utility of S-SAP markers was compared to that of AFLP markers. On the basis of our results, S-SAP is more informative marker system and showed higher average number of polymorphic bands per cultivar group than AFLP. Relationships among clones were analyzed by cluster analysis using unweighted pair-groups using arithmetic averages and in both cases revealed well defined groups of clones, in which ‘Malbec’ clones were separated from ‘Syrah’ clones. High variability of some clones could also be seen within these clusters. The different levels of polymorphism for ‘Malbec’ and ‘Syrah’ obtained in this study suggest that ‘Malbec’ exhibits a higher mutation frequency than ‘Syrah’. Our results indicate that higher proportion of polymorphic bands in S-SAP makes it a less labour-intensive and more efficient approach for developing markers for clonal identification.

Guidelines for the use and interpretation of assays for monitoring autophagy (4th edition)1.

In 2008, we published the first set of guidelines for standardizing research in autophagy. Since then, this topic has received increasing attention, and many scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Thus, it is important to formulate on a regular basis updated guidelines for monitoring autophagy in different organisms. Despite numerous reviews, there continues to be confusion regarding acceptable methods to evaluate autophagy, especially in multicellular eukaryotes. Here, we present a set of guidelines for investigators to select and interpret methods to examine autophagy and related processes, and for reviewers to provide realistic and reasonable critiques of reports that are focused on these processes. These guidelines are not meant to be a dogmatic set of rules, because the appropriateness of any assay largely depends on the question being asked and the system being used. Moreover, no individual assay is perfect for every situation, calling for the use of multiple techniques to properly monitor autophagy in each experimental setting. Finally, several core components of the autophagy machinery have been implicated in distinct autophagic processes (canonical and noncanonical autophagy), implying that genetic approaches to block autophagy should rely on targeting two or more autophagy-related genes that ideally participate in distinct steps of the pathway. Along similar lines, because multiple proteins involved in autophagy also regulate other cellular pathways including apoptosis, not all of them can be used as a specific marker for bona fide autophagic responses. Here, we critically discuss current methods of assessing autophagy and the information they can, or cannot, provide. Our ultimate goal is to encourage intellectual and technical innovation in the field

Curcumin potentiates the ErbB receptors inhibitor Afatinib for enhanced antitumor activity in malignant mesothelioma

several attempts have been made to develop targeted therapies for malignant mesothelioma (MM), an aggressive tumour with a poor prognosis. In this study we evaluated whether curcumin (CUR) potentiated the antitumor activity of the ErbB receptors inhibitor afatinib (AFA) on MM, employing cell lines cultured in vitro and mice bearing intraperitoneally transplanted, syngeneic MM cells. the rationale behind this hypothesis was that CUR could counteract mechanisms of acquired resistance to AFA. We analysed CUR and AFA effects on MM cell growth, cell cycle, autophagy, and on the modulation of tumour-supporting signalling pathways. this study demonstrated that, as compared to the individual compounds, the combination of AFA + CUR had a stronger effect on MM progression which can be ascribed either to increased tumour cell growth inhibition or to an enhanced pro-apoptotic effect. these results warrant future studies aimed at further exploring the therapeutic potential of AFA + CUR-based combination regimens for MM treatment

Pólen 2n e mecanismos meióticos de formação em Solanum commersonii ssp 2n pollen and meiotic mechanisms of Formation in Solanum commersonii ssp

Solanum commersonii subsp. commersonii Dun. e Solanum commersonii subsp. malmeanum Bitt. (2n=2x=24 - 1EBN) não podem ser diretamente cruzadas com a batata cultivada de S. tuberosum subsp. tuberosum L. (2n=4x=48 -4 EBN) em razão da barreira da ploidia e do número de equilíbrio do endosperma (EBN). No entanto, a produção de pólen não reduzido por S. commersonii ssp. (2x=2n=24 - 1EBN), permite o cruzamento desta com dihaplóides de batata e com as espécies diplóides S. phureja Juz. & Bukasov. e S. chacoense Bitt., que possuem 2 EBN. Conduziu-se este trabalho, com o objetivo de determinar o mecanismo meiótico e a freqüência dos grãos de pólen não reduzidos em cinco clones de Solanum commersonii ssp. Foram avaliados três clones de S. commersonii subsp. commersonii (SCC) e dois de S. commersonii subsp. malmeanum (SCM). Nas avaliações, duzentos grãos de pólen viáveis foram corados (carmim acético 2%) e mensurados, sendo considerados não reduzidos aqueles com o diâmetro igual ou superior a 25 µm. O clone Solanum commersonii subsp. commersonii Dun. (SCC 100) produz 26.1% de polen não reduzido, por fusos fundidos e fusos paralelos, o qual pode ser utilizado em programas de melhoramento.<br>Solanum commersonii subsp. commersonii Dun. and Solanum commersonii subsp. malmeanum Bitt. (2n=2x=24 -1EBN) cannot be directly crossed with the cultivated Solanum tuberosum subsp. Tuberosum L. (2n=4x=48 - 4EBN) species due to ploidy barriers and endosperm balance number (EBN). However, non-reduced pollen production by the wild species S. commersonii ssp. (2x=2n=24 -1EBN), allows the crossing of these species with dihaploids of potato and with the diploids species, S. phureja Juz. & Bukasov. and S. chacoense Bitt., all with 2EBN. The aim of this work was to determine the meiotic mechanism and frequency of non-reduced pollen production of five clones of S. commersonii ssp. Three different clones of S. commersonii subsp. commersonii Dun. (SCC) and two of S. commersonii subsp. malmeanum Bitt. (SCM) were evaluated. In the evaluation, two hundred viable pollen grains were stained (2% acetic carmin) and measured. The pollen grains measuring 25 µm in diameter or more were considered nonreduced. The clone Solanum commersonii subsp. commersonii Dun. (SCC 100) produces 26.1% of non-reduced pollen grains caused by parallel spindles and fusion spindles, which may be used in breeding program