The impact of immediate breast reconstruction on the time to delivery of adjuvant therapy: the iBRA-2 study

Background: Immediate breast reconstruction (IBR) is routinely offered to improve quality-of-life for women requiring mastectomy, but there are concerns that more complex surgery may delay adjuvant oncological treatments and compromise long-term outcomes. High-quality evidence is lacking. The iBRA-2 study aimed to investigate the impact of IBR on time to adjuvant therapy. Methods: Consecutive women undergoing mastectomy ± IBR for breast cancer July–December, 2016 were included. Patient demographics, operative, oncological and complication data were collected. Time from last definitive cancer surgery to first adjuvant treatment for patients undergoing mastectomy ± IBR were compared and risk factors associated with delays explored. Results: A total of 2540 patients were recruited from 76 centres; 1008 (39.7%) underwent IBR (implant-only [n = 675, 26.6%]; pedicled flaps [n = 105,4.1%] and free-flaps [n = 228, 8.9%]). Complications requiring re-admission or re-operation were significantly more common in patients undergoing IBR than those receiving mastectomy. Adjuvant chemotherapy or radiotherapy was required by 1235 (48.6%) patients. No clinically significant differences were seen in time to adjuvant therapy between patient groups but major complications irrespective of surgery received were significantly associated with treatment delays. Conclusions: IBR does not result in clinically significant delays to adjuvant therapy, but post-operative complications are associated with treatment delays. Strategies to minimise complications, including careful patient selection, are required to improve outcomes for patients

The “NeuroDante project”: neurometric measurements of subject’s reaction to literary auditory stimuli from Dante’s “Divina Commedia”

This work is a pilot study that used neurometric indexes during the listening of selected pieces of Dante‟s “Divina Commedia” in 20 participants. Half of them had a literary formation (Humanist; university students of litera-ture) while the other half of is attending other university courses (Not Human-ist). The study applied the electroencephalographic (EEG) rhythms variations, the heart rate (HR) and galvanic skin response (GSR) during the listening of the excerpts. The neurometric indexes here employed were the ApproachWith-drawal (AW), the cerebral Effort (CE) and the Emotional indexes (EI). Results for the comparisons of the estimated AW, CE and EI related to the perception of the canticas showed as the Humanist group reported higher AW and EI values when compared to the Not Humanist sample (p<0.03 and p<0.01, re-spectively). Results suggest that the perception of the aesthetic experience is significantly modulated by the previous specific knowledge experienced by the participants. Finally, results of this kind of research could find application in the implementation of software and devices based on symbiotic relation with the perspective reader or listener of a literature opera, in order to personalize and maximize the fruition of them

UMG Lenti: Novel Lentiviral Vectors for Efficient Transgene- and Reporter Gene Expression in Human Early Hematopoietic Progenitors

<div><p>Lentiviral vectors are widely used to investigate the biological properties of regulatory proteins and/or of leukaemia-associated oncogenes by stably enforcing their expression in hematopoietic stem and progenitor cells. In these studies it is critical to be able to monitor and/or sort the infected cells, typically via fluorescent proteins encoded by the modified viral genome. The most popular strategy to ensure co-expression of transgene and reporter gene is to insert between these cDNAs an IRES element, thus generating bi-cistronic mRNAs whose transcription is driven by a single promoter. However, while the product of the gene located upstream of the IRES is generally abundantly expressed, the translation of the downstream cDNA (typically encoding the reporter protein) is often inconsistent, which hinders the detection and the isolation of transduced cells. To overcome these limitations, we developed novel lentiviral dual-promoter vectors (named UMG-LV5 and –LV6) where transgene expression is driven by the potent UBC promoter and that of the reporter protein, EGFP, by the minimal regulatory element of the WASP gene. These vectors, harboring two distinct transgenes, were tested in a variety of human haematopoietic cell lines as well as in primary human CD34⁺ cells in comparison with the FUIGW vector that contains the expression cassette UBC-transgene-IRES-EGFP. In these experiments both UMG-LV5 and UMG–LV6 yielded moderately lower transgene expression than FUIGW, but dramatically higher levels of EGFP, thereby allowing the easy distinction between transduced and non-transduced cells. An additional construct was produced, in which the cDNA encoding the reporter protein is upstream, and the transgene downstream of the IRES sequence. This vector, named UMG-LV11, proved able to promote abundant expression of both transgene product and EGFP in all cells tested. The UMG-LVs represent therefore useful vectors for gene transfer-based studies in hematopoietic stem and progenitor cells, as well as in non-hematopoietic cells.</p></div

Schematic diagram of IRES-based and dual promoter lentiviral vectors.

<p>The expression cassettes of the lentiviruses used in this study are illustrated. The two IRES-containing vectors, FUIGW and UMG-LV11, differ for the position of the transgene and EGFP cDNA relative to the IRES element. In both viruses the transcription of this bicistronic unit is driven by the UBC promoter. The UMG-LV5 and UMG-LV6 vectors use independent promoters positioned back-to-back: UBC for the transgene and WASP (W) for EGFP. These dual-promoter vectors differ only for the orientation of the expression cassette. A short synthetic polyA signal, based on that of the human growth hormone gene, is downstream of the transcriptional unit in anti-sense orientation and is indicated by a diamond (♦).</p

Comparison of the transduction efficiency of FUIGW, UMG-LV5 and UMG-LV6 carrying the MSI2 cDNA in human hematopoietic cell lines.

<p>The cell lines K562, HL-60, MV4;11 and Jurkat were infected with FUIGW, UMG-LV5 or UMG-LV6 viruses carrying 3xFLAG-MSI2 cDNA as a transgene. As a control, void FUIGW vector was used. (<b>A</b>) Flow-cytometric analysis of EGFP expression in cells exposed to the relevant vectors. The percentages of EGFP-positive cells are indicated. (<b>B</b>) Whole-cell extracts, prepared as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0114795#s2" target="_blank">materials and methods</a>, were analyzed by Western blotting for FLAG-MSI2 and EGFP expression. Actin was used as a control for the amounts of extract loaded.</p

Efficiency of UMG-lenti vectors in the transduction of primary human CD34⁺ cells.

<p>CD34⁺ cells purified from cord blood were transduced with FUIGW, UMG-LV6 or UMG-LV11 viruses carrying the cDNAs for 3xFLAG-ZNF521 and EGFP. (<b>A</b>) FACS analysis of the transduced cells 5 days after transduction. The percentages of EGFP positive cells are indicated. (<b>B</b>) Western blotting analysis of FLAG-ZNF521 and EGFP expression was performed as described above on nuclear and cytosolic extracts. HDAC1 was used as a control for the amounts of extract loaded.</p