Protumorigenic effects of mir-145 loss in malignant pleural mesothelioma.

We identified a discrete number of microRNAs differentially expressed in benign or malignant mesothelial tissues. We focused on mir-145 whose levels were significantly downregulated in malignant mesothelial tissues and malignant pleural mesothelioma (MPM) cell lines as compared to benign tissues (pleura, peritoneum or cysts). We show that promoter hyper-methylation caused very low levels in MPM cell lines and specimens. Treatment of MPM cell lines with mir-145 agonists negatively modulated some protumorigenic properties of MPM cells, such as clonogenicity, cell migration and resistance to pemetrexed treatment. The main effector mechanism of the clonogenic death induced by mir-145 was that of accelerated senescence. We found that mir-145 targeted OCT4 via specific binding to its 3'-UTR. Increased intracellular levels of mir-145 decreased the levels of OCT4 and its target gene ZEB1, thereby counteracting the increase of OCT4 induced by pemetrexed treatment which is known to favor the development of chemoresistant cells. In line with this, reintroduction of OCT4 into mimic-145 treated cells counteracted the effects on clonogenicity and replicative senescence. This further supports the relevance of the mir-145-OCT4 interaction for the survival of MPM cells. The potential use of mir-145 expression levels to classify benign vs malignant mesothelial tissues and the differences between pemetrexed-induced senescence and that induced by the re-expression of mir-145 are discussed.

miR-204 targets Bcl-2 expression and enhances responsiveness of gastric cancer.

Micro RNAs (miRs) are small non-coding RNAs aberrantly expressed in human tumors. Here, we aim to identify miRs whose deregulated expression leads to the activation of oncogenic pathways in human gastric cancers (GCs). Thirty nine out of 123 tumoral and matched uninvolved peritumoral gastric specimens from three independent European subsets of patients were analyzed for the expression of 851 human miRs using Agilent Platform. The remaining 84 samples were used to validate miRs differentially expressed between tumoral and matched peritumoral specimens by qPCR. miR-204 falls into a group of eight miRs differentially expressed between tumoral and peritumoral samples. Downregulation of miR-204 has prognostic value and correlates with increased staining of Bcl-2 protein in tumoral specimens. Ectopic expression of miR-204 inhibited colony forming ability, migration and tumor engraftment of GC cells. miR-204 targeted Bcl-2 messenger RNA and increased responsiveness of GC cells to 5-fluorouracil and oxaliplatin treatment. Ectopic expression of Bcl-2 protein counteracted miR-204 pro-apoptotic activity in response to 5-fluorouracil. Altogether, these findings suggest that modulation of aberrant expression of miR-204, which in turn releases oncogenic Bcl-2 protein activity might hold promise for preventive and therapeutic strategies of GC.

Total upper and lower eyelid replacement following thermal burn using an ALT flap--a case report.

Upper and lower eyelid unilateral full thickness reconstruction in a patient with no available adjacent tissues because of burns or trauma sequelae is a surgical challenge. A case of severe thermal burn with unilateral complete defect of both upper and lower eyelids is reported, together with the surgical technique of reconstruction. The patient was a 65-year-old man who sustained deep burns of the head and neck with upper airway burns after falling into a fireplace. After tracheostomy and acute resuscitation, he underwent escharectomy and coverage of his head and neck burns with split thickness skin grafts and with full thickness skin grafts to the eyelids. There was incomplete take of the skin grafts to the upper and lower left eyelids. In these areas, infection and loss of the tarsum and subsequent eyelid retraction led to exposure keratitis and blurred vision. After healing and respiratory rehabilitation, he was referred to our microsurgical unit for upper and lower eyelid reconstruction. A free forearm flap was first considered, but the Allen test was negative. Therefore, a free anterolateral thigh (ALT) flap was chosen to provide skin eyelid coverage. The flap was harvested including fascia and centred on one perforator. The levator muscle stump and conjunctiva from both upper and lower cul-de-sacs were dissected and advanced. Flap vessels were anastomosed to the superficial temporal artery and vein. The conjunctiva and the fascia replaced the new inner upper and lower lamella. To our knowledge, this is the first report of the use of a perforator flap, the ALT flap, in full thickness reconstruction of both upper and lower eyelids and may be a reliable option in such selected and challenging situations.

Haemodynamic enhancement in perforator flaps: the inversion phenomenon and its clinical significance. A study of the relation of blood velocity and flow between pedicle and perforator vessels in perforator flaps.

Perforator flaps are perfused through a long vessel whose calibre decreases from its origin to the skin, because all branches have been sealed, resulting in a conduit with resistances in series, rather than a tree with resistances in parallel, as in the normal systemic circulation. This study was planned to assess whether the differences between perforator flap and normal systemic vasculature have an impact on haemodynamic parameters in perforator flaps and on their clinical significance. The study was performed on 10 patients. Echo-colour-Doppler measurement of diameters, velocity of flow and calculations of flow rate were made at the level of flap pedicle artery and skin perforator artery, pre- and post-operatively in each patient. Statistical analysis used the Wilcoxon matched pairs signed sum rank test. Our data show that in the donor area pre-operatively, blood velocity in skin artery perforator is lower that in the corresponding pedicle artery, whereas post-operatively, in perforator flaps, blood velocity in the perforator is higher than in the pedicle. The difference was statistically significant (P<0.01). There is an inversion of the gradient of blood velocity between pedicle artery and perforator artery compared to normal circulation. Furthermore, in normal circulation flow through the perforator was found smaller than that at the pedicle, whereas in perforator flaps, flow through the perforator is smaller but is a greater proportion of the flow through the pedicle and the difference is statistically significant (P<0.01). Therefore, the velocity of blood and the rate of flow reaching the skin are higher in perforator flaps than in normal circulation.