The Involvement of Epigenetic Mechanisms in HPV‐Induced Cervical Cancer

High‐risk human papillomavirus (HPV) genotypes infection associates with cervical dysplasia and carcinogenesis. hr‐HPV transforming potential is based on E6 and E7 viral oncoproteins actions on cellular proteins. A persistent infection with hr‐HPV leads to progression from precursor lesions to invasive cervical cancer inducing changes in host genome and epigenome. Pathogenesis and development of cancer associated with both genetic and epigenetic defects alter transcriptional program. An important role for malignant transformation in HPV‐induced cervical cancer is played by epigenetic changes that occur in both viral and host genome. Furthermore, there are observations demonstrating that oncogenic viruses, once they integrated into host genome, become susceptible to epigenetic alterations made by host machinery. Epigenetic regulation of viral gene expression is an important factor in HPV‐associated disease. Gene expression control is complex and involves epigenetic changes: DNA methylation, histone modification, and non‐coding RNAs activity. Persistent infection with hr‐HPV can cause viral DNA integration into host genome attracting defense mechanisms such as methylation machinery. In this chapter, we aim to review HPV infection role in chromatin modification/remodeling and the impact of HPV infection on non‐coding RNAs in cervix oncogenesis. The reversible nature of epigenetic alterations provides new opportunities in the development of therapeutic agents targeting epigenetic modification in oncogenesis

Biochemical Markers of Salt Stress in European Larch (Larix decidua)

[EN] Larix decidua, the European larch, does not grow in natural saline areas, but it can be affected by salinity either by the common practice of winter de-icing of mountain roads with NaCl, or when grown as an ornamental tree in urban areas by the use of low quality, salinised irrigation water. In the present study, the responses to salt stress of young seedlings obtained from seeds of seven Carpathian larch populations were analysed. After 30 days of treatments with 150 mM NaCl, growth and biochemical parameters were determinated. Salt stress inhibited growth of all seedlings, as shown by the relative reduction of stem length and fresh weight, and induced significant changes in the needle levels of several biochemical stress markers. Seedlings from all populations showed a marked reduction of photosynthetic pigments contents and an increase of proline and malondialdehyde (MDA) concentrations. Under salt stress, plants accumulated Na+ and Cl- in the needles, whereas K+ was maintained at a steady level. Responses of seedlings from the different populations were similar, with only small quantitative differences that did not allow the identification of more salt tolerant genotypes. However, the study revealed that several of the biochemical markers mentioned above can be suitable for the rapid and non-destructive assessment of the effects of salinity in European larch.Plesa, IM.; Al Hassan, M.; Sestras, A.; Vicente, O.; Boscaiu, M.; Sestras, R. (2018). Biochemical Markers of Salt Stress in European Larch (Larix decidua). Notulae Scientia Biologicae. 10(3):430-438. https://doi.org/10.15835/nsb10310322S43043810

Mechanisms of Oncogene Activation

The main modifications that characterize cancer are represented by alterations in oncogenes, tumor-suppressor genes, and non-coding RNA genes. Most of these alterations are somatic and the process is a multistep one. Tumors often arise from an initial transformed cell, and after subsequent genetic alterations different cytogenetically clones lead to tumor heterogeneity

Responses to Drought in Seedlings of European Larch (Larix decidua Mill.) from Several Carpathian Provenances

[EN] European larch (Larix decidua Mill.) has been reported either as more tolerant or as more sensitive to drought than conifers with perennial leaves. Previous studies have revealed that Carpathian populations of European larch display a high genetic variability. A comparative study of the responses of these populations to drought stress at the seedling stage might allow the identification of drought tolerant genotypes and reliable drought stress biomarkers, which could be eventually used for the early detection of drought effects in larch, not only under control greenhouse conditions, but also in their natural stands. Growth responses were analyzed in larch seedlings from six Romanian Carpathian populations, submitted to one month of mild drought stress under controlled conditions. Levels of photosynthetic pigments (chlorophylls a and b, and carotenoids), osmolytes (proline and total soluble sugars), monovalent cations (Na+ and K+), and malondialdehyde (MDA) and non-enzymatic antioxidants (total phenolics and flavonoids) were compared with control treatments and between populations. Growth and the pattern of the biochemical responses were very similar in the six populations. Drought stress lead to stem length decrease in all population, whereas reduction of fresh weight of needles was significant only in one population (BVVC), and reduction of water content of needles in two populations (BVVC and GuHo). The optimal biochemical traits for an early detection of drought symptoms in this species is the increase-in most populations-of total soluble sugars, MDA, and total phenolic compounds, whereas K+ reduction was significant in all populations. Photosynthetic pigments remained unchanged, except for the Anin population where they were reduced under stress. Multivariate principal component and hierarchical clustering analyses confirmed the impact of drought in the growth and physiology of European larch, and revealed that the humidity of the substrate was positively correlated with the growth parameters and the levels of K+ in needles, and negatively correlated with the levels of MDA, total soluble sugars, total phenolic compounds, and flavonoids in needles.Plesa, IM.; Al Hassan, M.; González-Orenga, S.; Sestras, A.; Vicente, O.; Prohens Tomás, J.; Boscaiu, M.... (2019). Responses to Drought in Seedlings of European Larch (Larix decidua Mill.) from Several Carpathian Provenances. Forests. 10(6):1-22. https://doi.org/10.3390/f10060511S122106Allen, C. D., Macalady, A. K., Chenchouni, H., Bachelet, D., McDowell, N., Vennetier, M., … Cobb, N. (2010). A global overview of drought and heat-induced tree mortality reveals emerging climate change risks for forests. 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Forest Ecology and Management, 261(4), 829-843. doi:10.1016/j.foreco.2010.10.038Bolte, A., Ammer, C., Löf, M., Madsen, P., Nabuurs, G.-J., Schall, P., … Rock, J. (2009). Adaptive forest management in central Europe: Climate change impacts, strategies and integrative concept. Scandinavian Journal of Forest Research, 24(6), 473-482. doi:10.1080/02827580903418224Xiang, W., Lei, X., & Zhang, X. (2016). Modelling tree recruitment in relation to climate and competition in semi-natural Larix-Picea-Abies forests in northeast China. Forest Ecology and Management, 382, 100-109. doi:10.1016/j.foreco.2016.09.050Sánchez-Gómez, D., Robson, T. M., Gascó, A., Gil-Pelegrín, E., & Aranda, I. (2013). Differences in the leaf functional traits of six beech (Fagus sylvatica L.) populations are reflected in their response to water limitation. Environmental and Experimental Botany, 87, 110-119. doi:10.1016/j.envexpbot.2012.09.011Bussotti, F., Pollastrini, M., Holland, V., & Brüggemann, W. (2015). Functional traits and adaptive capacity of European forests to climate change. Environmental and Experimental Botany, 111, 91-113. doi:10.1016/j.envexpbot.2014.11.006GRAMAZIO, P., PLESA, I. M., TRUTA, A. M., SESTRAS, A. F., VILANOVA, S., PLAZAS, M., … SESTRAS, R. E. (2018). Highly informative SSR genotyping reveals large genetic diversity and limited differentiation in European larch (Larixdecidua) populations from Romania. TURKISH JOURNAL OF AGRICULTURE AND FORESTRY, 42(3), 165-175. doi:10.3906/tar-1801-41Al Hassan, M., Chaura, J., López-Gresa, M. P., Borsai, O., Daniso, E., Donat-Torres, M. P., … Boscaiu, M. (2016). Native-Invasive Plants vs. Halophytes in Mediterranean Salt Marshes: Stress Tolerance Mechanisms in Two Related Species. Frontiers in Plant Science, 7. doi:10.3389/fpls.2016.00473Murray, F. W. (1967). On the Computation of Saturation Vapor Pressure. Journal of Applied Meteorology, 6(1), 203-204. doi:10.1175/1520-0450(1967)0062.0.co;2LICHTENTHALER, H. K., & WELLBURN, A. R. (1983). Determinations of total carotenoids and chlorophylls a and b of leaf extracts in different solvents. Biochemical Society Transactions, 11(5), 591-592. doi:10.1042/bst0110591Weimberg, R. (1987). Solute adjustments in leaves of two species of wheat at two different stages of growth in response to salinity. Physiologia Plantarum, 70(3), 381-388. doi:10.1111/j.1399-3054.1987.tb02832.xBates, L. S., Waldren, R. P., & Teare, I. D. (1973). Rapid determination of free proline for water-stress studies. Plant and Soil, 39(1), 205-207. doi:10.1007/bf00018060DuBois, M., Gilles, K. A., Hamilton, J. K., Rebers, P. A., & Smith, F. (1956). Colorimetric Method for Determination of Sugars and Related Substances. Analytical Chemistry, 28(3), 350-356. doi:10.1021/ac60111a017Hodges, D. M., DeLong, J. M., Forney, C. F., & Prange, R. K. (1999). Improving the thiobarbituric acid-reactive-substances assay for estimating lipid peroxidation in plant tissues containing anthocyanin and other interfering compounds. Planta, 207(4), 604-611. doi:10.1007/s004250050524Blainski, A., Lopes, G., & de Mello, J. (2013). Application and Analysis of the Folin Ciocalteu Method for the Determination of the Total Phenolic Content from Limonium Brasiliense L. Molecules, 18(6), 6852-6865. doi:10.3390/molecules18066852Zhishen, J., Mengcheng, T., & Jianming, W. (1999). The determination of flavonoid contents in mulberry and their scavenging effects on superoxide radicals. Food Chemistry, 64(4), 555-559. doi:10.1016/s0308-8146(98)00102-2Metsalu, T., & Vilo, J. (2015). ClustVis: a web tool for visualizing clustering of multivariate data using Principal Component Analysis and heatmap. Nucleic Acids Research, 43(W1), W566-W570. doi:10.1093/nar/gkv468Samaniego, L., Thober, S., Kumar, R., Wanders, N., Rakovec, O., Pan, M., … Marx, A. (2018). Anthropogenic warming exacerbates European soil moisture droughts. Nature Climate Change, 8(5), 421-426. doi:10.1038/s41558-018-0138-5Lindner, M., Fitzgerald, J. B., Zimmermann, N. E., Reyer, C., Delzon, S., van der Maaten, E., … Hanewinkel, M. (2014). Climate change and European forests: What do we know, what are the uncertainties, and what are the implications for forest management? Journal of Environmental Management, 146, 69-83. doi:10.1016/j.jenvman.2014.07.030Hlásny, T., Mátyás, C., Seidl, R., Kulla, L., Merganičová, K., Trombik, J., … Konôpka, B. (2014). Climate change increases the drought risk in Central European forests: What are the options for adaptation? Forestry Journal, 60(1), 5-18. doi:10.2478/forj-2014-0001Badalotti, A., Anfodillo, T., & Grace, J. (2000). Evidence of osmoregulation in Larix decidua at Alpine treeline and comparative responses to water availability of two co-occurring evergreen species. 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Vegetative desiccation tolerance: Is it a goldmine for bioengineering crops? Plant Science, 176(2), 187-199. doi:10.1016/j.plantsci.2008.10.002Corcuera, L., Gil-Pelegrin, E., & Notivol, E. (2011). Aridity promotes differences in proline and phytohormone levels in Pinus pinaster populations from contrasting environments. Trees, 26(3), 799-808. doi:10.1007/s00468-011-0651-xSchiop, S. T., Al Hassan, M., Sestras, A. F., Boscaiu, M., Sestras, R. E., & Vicente, O. (2015). Identification of Salt Stress Biomarkers in Romanian Carpathian Populations of Picea abies (L.) Karst. PLOS ONE, 10(8), e0135419. doi:10.1371/journal.pone.0135419Junker, L. V., & Ensminger, I. (2016). Relationship between leaf optical properties, chlorophyll fluorescence and pigment changes in senescingAcer saccharumleaves. Tree Physiology, 36(6), 694-711. doi:10.1093/treephys/tpv148FLEXAS, J. (2002). Drought-inhibition of Photosynthesis in C3 Plants: Stomatal and Non-stomatal Limitations Revisited. 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Interplay of Epigenetics with Gynecological Cancer

Recent data on the cell deregulation that occurs during the progression to cancer underlines the cooperation between genetic and epigenetic alterations leading to a malignant phenotype. Unlike genetic alterations, the epigenetic changes do not affect the DNA sequence of the genes, but determine the regulation of gene expression acting upon the genome. Moreover, unlike genetic changes, epigenetic ones are reversible, making them therapeutic targets in various conditions in general and in cancer disease in particular. The term epigenetics includes a series of covalent modifications that regulate the methylation pattern of DNA and posttranslational modifications of histones. Gene expression can also be regulated at the posttranscriptional level by microRNAs (miRNAs), a family of small noncoding RNAs that inhibit the translation of mRNA to protein. miRNAs can act as ‘oncomiRs’, as tumor suppressors, or both. In this chapter, we will (1) summarize the current literature on the key processes responsible for epigenetic regulation: DNA methylation, histone modifications and posttranscriptional gene regulation by miRNAs; (2) evaluate aberrant epigenetic modifications as essential players in cancer progression; (3) establish the roles of microenvironment-mediated epigenetic perturbations in the development of gynecological neoplasia; (4) evaluate epigenetic factors involved in drug resistance

Histone lysine demethylases as epigenetic modifiers in HPV-induced cervical neoplasia

Abstrac

Effects of Drought and Salinity on European Larch (Larix decidua Mill.) Seedlings

[EN] Larix decidua, the European larch, is not normally affected by drought or salinity in its natural habitats, but it may be when grown as an ornamental tree, by the widespread practice of winter de-icing of mountain roads with NaCl, and because of global warming-induced environmental changes. The responses of two-month-old larch seedlings to 30 days water deficit (withholding irrigation) or salt stress (150 mM NaCl) treatments were studied by determining stress-induced changes in several growth parameters and biochemical markers (ion and osmolyte contents, level of oxidative stress, activation of enzymatic and non-enzymatic antioxidant systems). Both treatments caused the inhibition of growth, degradation of photosynthetic pigments, a small increase in malondialdehyde (MDA, an oxidative stress biomarker), and the activation of antioxidant enzymes: superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), and glutathione reductase (GR). In all cases, salinity appeared to have stronger effects on the seedlings than water deficit. The presence of relatively high concentrations of glycine betaine, both in control and stressed plants, may represent a constitutive mechanism of defence against stress in European larch. Additionally, other responses were specific for salt stress and included the activation of K+ transport from roots to shoots and the accumulation of Pro as an osmoprotectantI.M.P. and M.A.H. were recipients of Erasmus Mundus pre-doctoral scholarships financed by the European Commission.Plesa, IM.; González-Orenga, S.; Al Hassan, M.; Sestras, AF.; Vicente, O.; Prohens Tomás, J.; Sestras, RE.... (2018). Effects of Drought and Salinity on European Larch (Larix decidua Mill.) Seedlings. Forests. 9(6). doi:10.3390/f9060320S9

Highly informative SSR genotyping reveals large genetic diversity and limited differentiation in European larch (Larix decidua) populations from Romania

[EN] European larch (Larix decidua) is naturally distributed at high elevations in Central Europe. Romanian populations of L. decidua are scattered in different areas of the country. In this study, we used 12 highly informative genomic simple sequence repeat (SSR) markers for genotyping seven populations from different areas of Romania. The SSR markers were highly variable, with up to 11 alleles per SSR locus and an average polymorphic information content of 0.713. High values of observed (Ho = 0.542) and expected (He = 0.738) heterozygosities were observed. Cluster analysis of populations did not group them according to geographical distance, but some clusters contained populations from a similar altitudinal range. The partition of genetic variation revealed that two-thirds of the genetic variation was found within individuals (due to high Ho levels), while only one-sixth of the total genetic variation corresponded to variation among populations. A population structure analysis identified four genetic clusters, and in most cases individuals from a single population were assigned to several genetic clusters. Multivariate principal coordinates analysis confirmed the population structure analysis. SSR markers are a powerful tool for evaluating diversity, relationships, and the genetic structure of Romanian L. decidua populations, which have high levels of diversity and low genetic structurationPietro Gramazio is grateful to Universitat Politecnica de Valencia for a predoctoral contract (Programa FPI de la UPV-Subprograma 1/2013 call). Ioana M Plesa is grateful to the Doctoral School of the University of Agricultural Sciences and Veterinary Medicine Cluj-Napoca and ICHAT - Institute of Advanced Horticulture Research of Transylvania, and Universitat Politecnica de Valencia, for their support. Mariola Plazas is grateful to Spanish Ministerio de Economia, Industria y Competitividad for a post doctoral grant within the Juan de la Cierva programme (FCJI-2015-24835).Gramazio, P.; Plesa, IM.; Truta, AM.; Sestras, AF.; Vilanova Navarro, S.; Plazas Ávila, MDLO.; Vicente, O.... (2018). Highly informative SSR genotyping reveals large genetic diversity and limited differentiation in European larch (Larix decidua) populations from Romania. Turkish Journal of Agriculture and Forestry. 42(3):165-175. https://doi.org/10.3906/tar-1801-41S16517542