Expression profiling of extramedullary acute myeloid leukemia suggests involvement of epithelial-mesenchymal transition pathways.

Extramedullary (EM) colonization is a rare complication of acute myeloid leukemia (AML), occurring in about 10% of patients, but the processes underlying tissue invasion are not entirely characterized. Through the application of RNAseq technology, we examined the transcriptome profile of 13 AMLs, 9 of whom presented an EM localization. Our analysis revealed significant deregulation within the extracellular matrix (ECM)-receptor interaction and focal-adhesion pathways, specifically in the EM sites. The transcription factor TWIST1, which is known to impact on cancer invasion by dysregulating epithelial-mesenchymal-transition (EMT) processes, was significantly upregulated in EM-AML. To test the functional impact of TWIST1 overexpression, we treated OCI-AML3s with TWIST1-siRNA or metformin, a drug known to inhibit tumor progression in cancer models. After 48 h, we showed downregulation of TWIST1, and of the EMT-related genes FN1 and SNAI2. This was associated with significant impairment of migration and invasion processes by Boyden chamber assays. Our study shed light on the molecular mechanisms associated with EM tissue invasion in AML, and on the ability of metformin to interfere with key players of this process. TWIST1 may configure as candidate marker of EM-AML progression, and inhibition of EMT-pathways may represent an innovative therapeutic intervention to prevent or treat this complication.

Circulating regulatory T cells in "clinical" monoclonal B-cell lymphocytosis.

Regulatory T-cells (Tregs) constitute a small subset of cells involved in antitumour immunity and are generally increased in patients with chronic lymphocytic leukemia (CLL). No data is available on Tregs in monoclonal B-cell lymphocytosis (MBL), a disease entity characterized by less than 5000/microL circulating clonal B-cells in absence of other features of lymphoproliferative disorders. We used multicolour flow cytometry to evaluate the number of circulating Tregs in 56 patients with "clinical" MBL, 74 patients with previously untreated CLL and 40 healthy subjects. MBL patients showed a lower absolute number of Tregs, compared to CLL patients, but slightly higher than controls. Moreover, the absolute cell number of Tregs directly correlated both with more advanced Rai/Binet clinical stages and peripheral blood B-cell lymphocytosis. Of note, the absolute number of Tregs was found lower in MBL patients than in CLL patients staged as 0/A Rai/Binet. The study showed that Treg increase gradually from normal subjects to "clinical" MBL patients and are significantly higher in CLL patients as compared to MBL patients. Moreover, a significant direct relationship was found between higher Treg values and a higher tumor burden expressed by B-lymphocytosis or more advanced clinical stages. In light of this data, MBL seems to be a preliminary phase preceding CLL. The progressive increase of Treg numbers might contribute both to the clinical evolution of MBL to overt CLL and to CLL progression.

Effects of prolonged high-altitude exposure on peripheral adrenergic receptors in young healthy volunteers.

The regulation of adrenergic receptors during hypoxia is complex, and the results of published reports have not been consistent. In the present study blood cell adrenoceptor characteristics at sea level (SL) before and after prolonged exposure to high altitude (HA) were measured in seven trained young male lowlanders. Sympathoadrenal activity and clinical haemodynamic parameters were also evaluated before departure (SLB), after 1 week (HA1) and 4 weeks (HA4) at HA and 1 week after return to sea level (SLA). As compared to pre-departure sea level values, urinary norepinephrine excretion increased significantly during altitude exposure [SLB: 10.26 (3.04) microg x 3 h(-1); HA1: 23.2 (4.19) microg x 3 h(-1); HA4: 20.3 (8.68) microg x 3 h(-1)] and fell to pre-ascent values 1 week after return to sea level [SLA: 9 (2.91) microg x 3 h(-1)]. In contrast, mean urinary epinephrine levels did not increase over time at HA. Both systolic and diastolic blood pressures, as well as heart rate, were increased during HA exposure. The circadian blood pressure and heart rate rhythms were preserved during all phases of altitude exposure. Mean maximal binding (Bmax) of the alpha2-adrenoceptor antagonist [3H]rauwolscine to platelet membranes was significantly reduced (P < 0.001) after exposure to chronic hypoxia [SLB: 172.6 (48.5) fmol x mg(-1) protein versus SLA: 136.8 (56.1) fmol x mg(-1) protein] without change in the dissociation constant (K(D)). Neither the lymphomonocyte beta2-adrenoceptor Bmax [SLB: 38.5 (13.6) fmol x mg(-1) protein, versus SLA: 32.4 (12.1) fmol mg(-1) protein] nor the K(D) for [3H]dihydroalprenolol was affected by chronic hypoxia. Cyclic AMP (adenosine 3',5'-cyclic monophoshate) generation in lymphomonocytes by maximal isoproterenol stimulation was not modified after prolonged HA exposure. In conclusion, the down-regulation of alpha2-adrenoceptors appears to be an important component of the adrenergic system response to HA exposure.

Effects of central alpha 2-adrenergic agonists on systemic haemodynamics and on baroreceptor reflex sensitivity in spontaneously hypertensive rats.

Clonidine, an imidazoline derivative, is a widely used antihypertensive agent which acts by stimulating the alpha 2-adrenergic receptors at the central nervous system. Clonidine also seems to exert beneficial effects on baroreceptor reflex sensitivity which is usually altered by arterial systemic hypertension and arteriosclerosis. Aim of the study was to compare acute and chronic hypotensive response and the effects on baroreflex sensitivity of a newly synthesized central alpha 2-adrenergic agonist, guanfacine, with those induced by clonidine. The effects of acute and chronic treatment were studied by evaluating blood pressure modifications through intraarterial or sphygmomanometric detection. Baroreflex sensitivity was studied before and after acute treatment with the two drugs. Our results show that guanfacine and clonidine may induce comparable effects, both reducing arterial blood pressure levels and heart rate with an order of potency for guanfacine 10 times lower than clonidine. After chronic treatment the effect of guanfacine on blood pressure was more pronounced than that observed after clonidine administration. Both drugs may enhance baroreflex sensitivity, with a more evident effect of guanfacine than clonidine in response to a sudden decrease of blood pressure. This suggests that guanfacine could afford a more effective cardiovascular adaptation during acute hypotension (i.e. during orthostatic hypotension), particularly in the elderly hypertensive patients.