Are early somatic embryos of the norway spruce (Picea abies (L.) Karst.) organised?

Background Somatic embryogenesis in conifer species has great potential for the forestry industry. Hence, a number of methods have been developed for their efficient and rapid propagation through somatic embryogenesis. Although information is available regarding the previous process-mediated generation of embryogenic cells to form somatic embryos, there is a dearth of information in the literature on the detailed structure of these clusters. Methodology/Principal Findings The main aim of this study was to provide a more detailed structure of the embryogenic tissue clusters obtained through the in vitro propagation of the Norway spruce (Picea abies (L.) Karst.). We primarily focused on the growth of early somatic embryos (ESEs). The data on ESE growth suggested that there may be clear distinctions between their inner and outer regions. Therefore, we selected ESEs collected on the 56th day after sub-cultivation to dissect the homogeneity of the ESE clusters. Two colourimetric assays (acetocarmine and fluorescein diacetate/propidium iodide staining) and one metabolic assay based on the use of 2,3,5-triphenyltetrazolium chloride uncovered large differences in the metabolic activity inside the cluster. Next, we performed nuclear magnetic resonance measurements. The ESE cluster seemed to be compactly aggregated during the first four weeks of cultivation; thereafter, the difference between the 1H nuclei concentration in the inner and outer clusters was more evident. There were clear differences in the visual appearance of embryos from the outer and inner regions. Finally, a cluster was divided into six parts (three each from the inner and the outer regions of the embryo) to determine their growth and viability. The innermost embryos (centripetally towards the cluster centre) could grow after sub-cultivation but exhibited the slowest rate and required the longest time to reach the common growth rate. To confirm our hypothesis on the organisation of the ESE cluster, we investigated the effect of cluster orientation on the cultivation medium and the influence of the change of the cluster’s three-dimensional orientation on its development. Maintaining the same position when transferring ESEs into new cultivation medium seemed to be necessary because changes in the orientation significantly affected ESE growth. Conclusions and Significance This work illustrated the possible inner organisation of ESEs. The outer layer of ESEs is formed by individual somatic embryos with high metabolic activity (and with high demands for nutrients, oxygen and water), while an embryonal group is directed outside of the ESE cluster. Somatic embryos with depressed metabolic activity were localised in the inner regions, where these embryonic tissues probably have a very important transport function

Development of a T Cell Receptor Targeting an HLA-A*0201 Restricted Epitope from the Cancer-Testis Antigen SSX2 for Adoptive Immunotherapy of Cancer.

The clinical success of adoptive immunotherapy of cancer relies on the selection of target antigens that are highly expressed in tumor cells but absent in essential normal tissues. A group of genes that encode the cancer/testis or cancer germline antigens have been proposed as ideal targets for immunotherapy due to their high expression in multiple cancer types and their restricted expression in immunoprivileged normal tissues. In the present work we report the isolation and characterization of human T cell receptors (TCRs) with specificity for synovial sarcoma X breakpoint 2 (SSX2), a cancer/testis antigen expressed in melanoma, prostate cancer, lymphoma, multiple myeloma and pancreatic cancer, among other tumors. We isolated seven HLA-A2 restricted T cell receptors from natural T cell clones derived from tumor-infiltrated lymph nodes of two SSX2-seropositive melanoma patients, and selected four TCRs for cloning into retroviral vectors. Peripheral blood lymphocytes (PBL) transduced with three of four SSX2 TCRs showed SSX241-49 (KASEKIFYV) peptide specific reactivity, tumor cell recognition and tetramer binding. One of these, TCR-5, exhibited tetramer binding in both CD4 and CD8 cells and was selected for further studies. Antigen-specific and HLA-A*0201-restricted interferon-γ release, cell lysis and lymphocyte proliferation was observed following culture of TCR engineered human PBL with relevant tumor cell lines. Codon optimization was found to increase TCR-5 expression in transduced T cells, and this construct has been selected for development of clinical grade viral vector producing cells. The tumor-specific pattern of expression of SSX2, along with the potent and selective activity of TCR-5, makes this TCR an attractive candidate for potential TCR gene therapy to treat multiple cancer histologies

Scented grasses in Norway - Identity and uses

Published version. Source at http://doi.org/10.1186/s13002-015-0070-y.Background: Some grass species are richer in coumarin and thus more sweetly scented than others. These have been eagerly sought after in parts of Norway, but the tradition has been weakly documented, both in terms of the species collected, their vernacular names, and uses. Methods: Based on literature data and a substantial body of information collected during my own ethnobotanical field work, artefacts and voucher specimens, the grass species are identified, and their uses clarified. Results: In Norwegian literature, the tradition of collecting and using scented grasses has received little attention, and past authors largely refer it to Anthoxanthum spp. The tradition’s concentration to the Sámi strongholds of northernmost Norway, and most authors’ lacking knowledge of the Sámi language, have contributed to the weak and misleading coverage in previous publications. Coumarin-rich grass species are well known in folk tradition in northernmost Norway, as luktegress (Norwegian, “scent grass”), háissasuoidni (North Sámi, “scent grass”), hajuheinä (Finnish, “scent grass”), or similar terms. They have been (and still are) frequently collected, and used as perfume, for storing with clothes, and a number of other purposes. Despite literature records identifying the species used as Anthoxanthum odoratum coll. (including A. nipponicum), the main source utilized in North Norway is Hierochloë odorata, both ssp. arctica and ssp. odorata. Anthoxanthum nipponicum and Milium effusum are alternative, but infrequently used sources of material, depending on local tradition and availability. Conclusion: By far the most important grass species hiding behind the “scented grass” tradition in Norway is Hierochloë odorata. Anthoxanthum nipponicum is also used, but much less frequently, and only a single record confirms the use of Milium effusum. Only the foliage of Hierochloë provides suitable material for making traditional braids. The three major ethnic groups in Norway have all utilized scented grasses as perfume and for storing with clothes, but the tradition’s geographical concentration to the far north of Norway (Finnmark and NE Troms), suggests that it has originally mainly been a Sámi tradition, adopted by their neighbours