Tumor necrosis is associated with increased alphavbeta3 integrin expression and poor prognosis in nodular cutaneous melanomas

<p>Abstract</p> <p>Background</p> <p>Tumor necrosis and apoptotic activity are considered important in cancer progression, but these features have not been much studied in melanomas. Our hypothesis was that rapid growth in cutaneous melanomas of the vertical growth phase might lead to tissue hypoxia, alterations in apoptotic activity and tumor necrosis. We proposed that these tumor characteristics might be associated with changes in expression of cell adhesion proteins leading to increased invasive capacity and reduced patient survival.</p> <p>Methods</p> <p>A well characterized series of nodular melanoma (originally 202 cases) and other benign and malignant melanocytic tumors (109 cases) were examined for the presence of necrosis, apoptotic activity (TUNEL assay), immunohistochemical expression of hypoxia markers (HIF-1 α, CAIX, TNF-α, Apaf-1) and cell adhesion proteins (α_vβ₃integrin, CD44/HCAM and osteopontin). We hypothesized that tumor hypoxia and necrosis might be associated with increased invasiveness in melanoma through alterations of tumor cell adhesion proteins.</p> <p>Results</p> <p>Necrosis was present in 29% of nodular melanomas and was associated with increased tumor thickness, tumor ulceration, vascular invasion, higher tumor proliferation and apoptotic index, increased expression of α_vβ₃integrin and poor patient outcome by multivariate analysis. Tumor cell apoptosis did also correlate with reduced patient survival. Expression of TNF-α and Apaf-1 was significantly associated with tumor thickness, and osteopontin expression correlated with increased tumor cell proliferation (Ki-67).</p> <p>Conclusion</p> <p>Tumor necrosis and apoptotic activity are important features of melanoma progression and prognosis, at least partly through alterations in cell adhesion molecules such as increased α_vβ₃integrin expression, revealing potentially important targets for new therapeutic approaches to be further explored.</p

Interactive and Single Effects of Ectomycorrhiza Formation and Bacillus cereus on Metallothionein MT1 Expression and Phytoextraction of Cd and Zn by Willows

Single and joint ectomycorrhizal (+ Hebeloma mesophaeum) and bacterial (+ Bacillus cereus) inoculations of willows (Salix viminalis) were investigated for their potential and mode of action in the promotion of cadmium (Cd) and zinc (Zn) phytoextraction. Dual fungal and bacterial inoculations promoted the biomass production of willows in contaminated soil. Single inoculations either had no effect on the plant growth or inhibited it. All inoculated willows showed increased concentrations of nutritional elements (N, P, K and Zn) and decreased concentrations of Cd in the shoots. The lowest biomass production and concentration of Cd in the willows (+ B. cereus) were combined with the strongest expression of metallothioneins. It seems that biotic stress from bacterial invasion increased the synthesis of these stress proteins, which responded in decreased Cd concentrations. Contents of Cd and Zn in the stems of willows were combination-specific, but were always increased in dual inoculated plants. In conclusion, single inoculations with former mycorrhiza-associated B. cereus strains decreased the phytoextraction efficiency of willows by causing biotic stress. However, their joint inoculation with an ectomycorrhizal fungus is a very promising method for promoting the phytoextraction of Cd and Zn through combined physiological effects on the plant

EPIP as an abscission promoting agent in the phytohormonal pathway

Understanding the mechanisms underlying the activation of the abscission zone (AZ) responsible for organ separation from plant body in crop species will help improve their yielding and economic importance. Special attention has been given recently to the role of the INFLORESCENCE DEFICIENT IN ABSCISSION protein, particularly its functional fragment, EPIP peptide. Its stimulatory effect on abscission in different crops has been demonstrated. Recently we described the role of EPIP in the redox, lipid, and pectin-related events taking place in AZ of Lupinus luteus flowers, which undergo massive abscission in natural conditions. To further examine EPIP contribution in AZ functioning, here, we analyze its impact on the ultrastructural changes, synthesis of two hormonal abscission stimulators - abscisic acid (ABA) and ethylene (ET), and the appearance of phosphoproteins. As our results show, the response of flower AZ to exogenous EPIP involves the induction of distinct modifications related to the one hand with upregulation of cell activity but on the other hand degradation processes and possible autophagy. Furthermore, the EPIP stimulated biosynthesis pathways of ABA and ET precisely in AZ cells. In addition, progressive phosphorylation of proteins has been observed under EPIP influence. The highly accumulated ones were identified as those, related to primary metabolism and reactive oxygen species homeostasis, and their role in abscission has been discussed. To summarizing, the presented detailed description of EPIP action in AZ cells in combination with our previous data offers new insights into its regulatory function and provides opportunities to counteract excessive flower abscission in lupine

Transcribed DNA is preferentially located in the perichromatin region of mammalian cell nuclei

The precise localization of transcribed DNA and resulting RNA is an important aspect of the functional architecture of the nucleus. To this end we have developed a novel in situ hybridization approach in combination with immunoelectron microscopy, using sense and anti-sense RNA probes that are derived from total cellular or cytoplasmic poly(A+) RNA. This new technology is much more gentle than classical in situ hybridization using DNA probes and shows excellent preservation of nuclear structure. Carried out on ultrathin sections of fixed and resin-embedded COS-7 cells, it revealed at high resolution the localization of the genes that code for the cellular mRNAs. Quantitative analysis shows that most transcribed DNA is concentrated in the perichromatin region, i.e. the interface between subchromosomal compact chromatin domains and the interchromatin space essentially devoid of DNA. The RNA that is produced is found mainly in the perichromatin region and the interchromatin space. These results imply that in the mammalian nucleus the chromatin fiber is folded so that active genes are predominantly present in the perichromatin region, which is the most prominent site of transcription